Category Archives: Stem Cell Medical Center

In situ repair abilities of human umbilical cordderived mesenchymal stem cells and autocrosslinked hyaluronic acid gel complex in rhesus monkeys with…

Abstract

Increasing occurrence of moderate to severe intrauterine adhesion (IUA) is seriously affecting the quality of human life. The aim of the study was to establish IUA models in nonhuman primates and to explore the dual repair effects of human umbilical cordderived mesenchymal stem cells (huMSCs) loaded on autocrosslinked hyaluronic acid gel (HA-GEL) on endometrial damage and adhesion. Here, we recorded the menstrual cycle data in detail with uterine cavities observed and endometrial tissues detected after intervention, and the thicker endometria, decreased amount of fibrotic formation, increased number of endometrium glands, etc., suggested that both HA-GEL and huMSC/HA-GEL complexes could partially repair IUA caused by mechanical injury, but huMSC/HA-GEL complex transplantation had notable dual repair effects: a reliable antiadhesion property and the promotion of endometrial regeneration.

Intrauterine adhesion (IUA), known as Asherman syndrome, is described as the partial or complete binding of the uterine cavity due to the buildup of scar tissue formation in the upper functional layer, resulting from endometrial damage to the lower basal layer (1). Repeated intrauterine operations, such as dilatation and curettage (D&C) and hysteroscopy, are the main cause of the common prevalence of IUA, with approximately 45% of moderate-to-severe cases occurring in these circumstances (2). Severe endometrium dysfunction will cause women within reproductive age to have this reproductive disadvantage, which was once considered a terminal disease that caused infertility (3, 4).

Therefore, the vital aim for IUA treatment is to re-establish the uterine cavity and restore endometrial function. The current and standard operation method for IUA is hysteroscopic transcervical resection of adhesion (TCRA), while the preferred alternative involves the combined application of various adjuvant therapies, including physical barriers (contraceptive device, intrauterine balloon device, Foley balloon, etc.) for postoperative adhesion and estrogen therapy and amnion graft for endometrial regeneration (58). Although there is a certain therapeutic efficacy of these antiadhesion strategies, there are some disadvantages and shortcomings to the strategies that cannot be ignored, such as resistance to secondary surgery, limited area of isolation, induction of intrauterine inflammatory response, and difficulty in endometrial regeneration (9). Further, the high recurrence rate of postoperative adhesion and the low clinical pregnancy rate are still a focus and are universally recognized as a problem for patients with severe IUAs (4, 10).

Autocrosslinked hyaluronic acid gel (HA-GEL), another promising physical barrier with natural mix of extracellular matrix and synovial fluid, has been approved by China Food & Drug Administration (CFDA) as a medical device for clinical practice after hysteroscopic adhesiolysis to achieve improvement in histocompatibility and viscosity, and the American Association of Gynecologic Laparoscopists has reported the effectiveness of HA-GEL in the prevention of IUAs in 2017 (11). The application of HA-GEL in uterine cavity treatment has its own advantages of few degradations by product retention following the outflow of menstrual blood (11, 12). Compared to the previously used biomaterials, HA-GEL has a prolonged absorption time (as long as 7 to 14 days), the expansion characteristics of which can continuously isolate the postoperative uterine cavity to resist adhesion recurrence, and its other material properties can regulate the inflammatory response and repair endometrial injuries. Related experimental and clinical studies have suggested that HA-GEL is effective in the prevention of postoperative adhesion, and when combined application with a TCRA operation, it showed increases (~58.5%) in the effective rate of postoperative antiadhesion and decreases (~66.1%) in the postoperative recurrence rate (13). However, for severe IUA with a seriously injured basal layer and a loss of functional endometrium, endometrial regeneration remains an enormous challenge owing to the limited efficacy of current interventions.

Recently, stem cellbased therapy has emerged as a promising and exciting method of tissue regeneration (1416). Human umbilical cordderived mesenchymal stem cells (huMSCs) originate from the embryonic mesoderm and have the potential for multipotent differentiation; they have been regarded as a promising and extensive source for cell-based therapies due to their easy collection from discarded umbilical cords and their low immunogenicity. Some studies have shown the potential of huMSCs to repair damaged tissue (1720), and the feasibility of stem cells in restoring the endometrial structure and function has also been verified by additional clinical and experimental studies (2124). In this study, we used rhesus monkeys to construct a previously unidentified animal model of IUA and aimed to develop a complex of huMSCs loaded on HA-GEL to increase the local perseverance and activity of the stem cells and to improve the poor prognosis with the following dual functions: preventing postoperative adhesion with biomaterials and repairing the full layer of uterine wall. We also aimed to analyze the related repair or endometrial injuries, to study the motivation behind endometrial regeneration, and to explore the underlying mechanisms.

huMSCs [passage 3 (P3) to P9] had an appearance that was similar to typical spindle-shaped fibroblast-like cells, and they were arranged closely with vortex-like growth (fig. S1A1). The positive cells that were expanded in the enriching culture were successfully induced to become osteoblasts with bone matrix formation and adipocytes with lipid droplet formation (fig. S1A2 and A3). In addition, fluorescence-activated cell sorting (FACS) showed that the targeted cells expressed CD44 (99.40%), CD73 (99.56%), CD90 (99.92%), and CD105 (99.80%), but not CD34, CD45, and HLA-DR (<1%; fig. S1B).

To further evaluate the safety of huMSCs on HA-GEL, FACS results preliminarily verified that there was a low parentage of apoptotic cells in the coculture group (huMSCs/HA-GEL), and there was no notable difference between the coculture group and the culture-separated group (huMSCs) (fig. S2, A to C). In addition, the live-dead cell staining result we obtained before was added, and the result showed a small number of dead cells in both the huMSCs/HA-GEL and huMSCs groups, without significant difference compared with that in the culture-separated group (huMSCs) (fig. S2, D and E).

Two months after endometrial intervention by uterine D&C, all six monkeys stopped menstruating; smaller uterine cavities and pale and uneven endometrial surfaces were observed, which had an adhesive zone full of endometrial cavity fluid. Thinner endometrial tissue was detected and observed under Doppler ultrasound scanning with discontinuous endometria and strong echo (Fig. 1A), and the endometrial thickness (1.9833 0.4298 mm) after mechanical injury showed significant differences when compared with the thickness (4.0333 0.5185 mm) before intervention (P < 0.01, n = 6; Fig. 1B and table S1). Changes in the structure of the endometrial tissues were assessed by hematoxylin and eosin (H&E) staining. Two months after mechanical injury, the endometrium was disorganized and had few or no glands (Fig. 1C). Endometrial gland numbers decreased markedly compared with those of the premechanical injury (0.6839 0.8608 versus 6.8576 2.6901 per unit area, respectively) (P < 0.001, n = 6; Fig. 1D and table S1). Similarly, to further evaluate the degree of fibrosis, Masson staining was performed at 2 months after mechanical injury (Fig. 1E). Increased fibrotic area ratios were detected and were analyzed quantitatively; more collagen deposition was observed at 2 months after mechanical injury compared with that of the premechanical injury (0.6557 0.6359% versus 0.0716 0.0942%) (P < 0.05, n = 6; Fig. 1F and table S1).

(A) Detection of Doppler ultrasound. A1: Representative image of endometrial thickness for pre-D&C; A2: Representative image of endometrial thickness at 2 months post-D&C (red arrow, the endometrium echo; blue area, the largest cross section of endometrium). (B) Comparisons of endometrial thickness for pre- or post-D&C. (C) H&E staining of endometria for pre-D&C (C1, C3, and C5) and post-D&C (C2, C4, and C6); 10401, 10403, and 10406, respectively; see table S3 for details. Inserted overview pictures are of lower magnification; black squares are highly magnified regions. (D) Masson staining of endometria for pre-D&C (D1, D3, and D5) and post-D&C (D2, D4, and D6); 10401, 10403, and 10406, respectively; see table S3 for details. Inserted overview pictures are of lower magnification; black squares are highly magnified regions. (E) Comparisons of endometrial gland numbers per unit area for pre- or post-D&C. (F) Comparisons of fibrotic area ratios for pre- or post-D&C. *P < 0.05, **P < 0.01, and ***P < 0.001 versus the pre-D&C group, and the results shown are the mean SEM of three technical replicates from each animal.

Two months after the huMSCs/HA-GEL complex was transplanted into the uterine cavity, menstruation resumed cycling in all monkeys, and there were significantly more endometrial gland numbers (4.9662 1.4935, per unit area) than there were (3.6320 1.0060, per unit area) after HA-GEL transplantation alone (P < 0.01; Fig. 2, A and B, and table S2). Moreover, the huMSCs/HA-GEL transplantation group showed marked decreases in fibrotic areas (5.5955 3.6572%) compared with that of the HA-GEL transplantation group (14.2131 13.7193%) (P < 0.01; Fig. 2, C and D, and table S2).

(A) Endometrial H&E staining at 2 months after HA-GEL transplantation (A1, A3, and A4 correspond to 10401, 10403, and 10404, respectively) and huMSCs/HA-GEL transplantation (A2, A5, and A6 correspond to 10402, 10405, and 10406, respectively); 10401 to 10406, see table S3 for details. (B) Endometrial Masson staining at 2 months after HA-GEL transplantation (B1, B3, and B4 correspond to 10401, 10403, and 10404, respectively) and huMSCs/HA-GEL transplantation (B2, B5, and B6 correspond to 10402, 10405, and 10406, respectively); 10401 to 10406, see table S3 for details. (C) Comparisons of endometrial gland numbers per unit area between the HA-GEL transplantation group and the huMSC/HA-GEL transplantation group. (D) Comparisons of fibrotic area ratios between the HA-GEL transplantation group and the huMSCs/HA-GEL transplantation group. ##P < 0.01 versus HA-GEL transplantation group, and the results shown are the mean SEM of three technical replicates from each animal.

Abdominal surgeries were carried out, and three normal uterine cavities were exposed and revealed a thicker endometrium without an adhesive zone and endometrial cavity fluid in the huMSCs/HA-GEL transplantation group, whereas three uterine cavities in the HA-GEL transplantation group were still found to be abnormal with a mild to moderate amount of adhesion and a thinner rough endometrium (Fig. 3A). In addition, the smooth and thicker endometrial tissue with a third-line echo was also revealed and verified by ultrasound examination in the huMSCs/HA-GEL transplantation group (Fig. 3B), and the endometrial thickness (4.2667 0.5558 mm) was significantly different compared with that (1.0667 0.6650 mm) in the HA-GEL transplantation group (P < 0.01; Fig. 3C and table S2). Furthermore, the ultrastructure of the endometrium in the huMSCs/HA-GEL transplantation group showed short and sparse microvilli on the surface of epithelial cells, mucinous secretions in the glandular cavity with orderly arranged cells, tight intercellular junctions, and obvious edema of stroma, but the endometrial ends were uneven and the cellular edges had a frayed morphology. Further, loose connections between cells were observed in the HA-GEL transplantation group (Fig. 3D).

(A) Representative images of uterine cavities in the HA-GEL transplantation and huMSCs/HA-GEL transplantation groups (the dotted area and the red arrow mark the endometrial area). (B) Representative images of endometrial thickness for ultrasound detection in the HA-GEL transplantation and huMSCs/HA-GEL transplantation groups (the red arrow marks the endometrial echo; the blue area marks the largest cross section of the endometrium). (C) Comparisons of endometrial thickness between the HA-GEL transplantation group and the huMSCs/HA-GEL transplantation group. (D) Representative images of ultrastructural changes in the HA-GEL transplantation and huMSCs/HA-GEL transplantation groups (the left panel shows the surface of epithelial cells; the right panel shows the intercellular changes). ##P < 0.01 versus 2 months postHA-GEL, and the results shown are the mean SEM of three technical replicates from each animal. Photos provided by Lingjuan Wang and Chengliang Xiong (Institute of Reproductive Health, Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology).

By systematic and comprehensive comparison of endometrial tissues before surgery, increased gland numbers were found both in the HA-GEL transplantation group (3.63 1.01 versus 0.68 0.86, respectively, per unit area; P < 0.001) and in the huMSCs/HA-GEL transplantation group (4.97 1.49 versus 0.68 0.86, respectively, per unit area; P < 0.001). The gland numbers were approaching normal levels (6.86 2.69, per unit area; pre-D&C) 2 months after huMSCs/HA-GEL transplantation (Fig. 4A and table S3). Conversely, the Masson staining showed an increasing degree of fibrotic aggravation 2 months after HA-GEL transplantation (14.21 13.72% versus 0.66 0.64%; P < 0.05), but there was only a slight increase in fibrosis and some relief of aggravation after transplantation of the huMSCs/HA-GEL complex (5.60 3.66 versus 0.66 0.64; P < 0.01) (Fig. 4B and table S3). There was no significant difference in endometrial thickness or after mechanical injury (1.07 0.67 versus 1.98 0.41 mm) 2 months after HA-GEL transplantation compared with that of the HA-GEL transplantation, while the endometrial thickness was notably increased after transplantation of the huMSCs/HA-GEL complex (4.27 0.56 versus 1.98 0.41 mm; P < 0.01) that was similar to the normal levels observed before mechanical injury of the endometria (4.03 0.52 mm; pre-D&C) (Fig. 4C and table S3).

(A) Comparisons of endometrial gland numbers per unit area. (B) Comparisons of ratios of fibrotic area (%). (C) Comparisons of endometrial thickness (mm). *P < 0.05, **P < 0.01, and ***P < 0.001, all versus pre-D&C; ###P < 0.001 versus pre-D&C; #P < 0.05, ##P < 0.01, and ###P < 0.001, all versus 2 months post-D&C; ##P < 0.01 versus 2 months postHA-GEL, and the results shown are the mean SEM of three technical replicates from each animal.

The probe Vysis SRY Probe LSI SRY Spectrum Orange/Vysis CEP X Spectrum Green was used to mark huMSCs by a fluorescence in situ hybridization (FISH) technique. However, it was unclear if there was homology of probe sequences for the Yp11.3 region [sex-determining region Y (SRY), associated probe sequence] and DXZ1 (Xp11.1-Xq11.1) (CEP X, associated probe sequence) between human and rhesus monkeys. By directly extracting DNA from huMSCs (containing XY chromosome) and the spleens of rhesus monkeys, it was verified that the two probe sequences did not share homology between human and rhesus monkey (fig. S3). Then, human endometrial tissue was obtained as a positive control group (Fig. 5A), and one of the three endometria in the HA-GEL transplantation group was randomly selected as a negative control (Fig. 5B). FISH detection showed the absence of a positive signal (green/orange double signal or green signal) in the endometria 2 months after huMSCs/HA-GEL transplantation (Fig. 5, C to E), suggesting that huMSCs failed to locate to the endometrium after transplantation into the uterine cavity.

(A) Positive control, human endometrium (containing XX chromosomes); the red arrow indicates the green signal for Vysis CEP X (DXZ1). (B) Endometrial localization of huMSCs in the HA-GEL transplantation group (negative control). (C to E) Distribution of huMSCs in endometria 2 months after huMSCs/HA-GEL co-transplantation. Double/single-labeled staining (orange/green signal or just green signal) cells were defined as huMSCs. For details on 10402, 10405, and 10406, see table S3. Inserted overview pictures show a lower magnification.

Furthermore, potential cytokines secreted by huMSCs were further detected in the endometria by immunofluorescence staining, and as expected, increased positive expression was found in the endometria of the huMSCs/HA-GEL transplantation group; there were significant differences in insulin-like growth factor (IGF-1), epidermal growth factor (EGF), and brain-derived neurotrophic factor (BDNF) between the two transplanted groups (P < 0.05), but there were no significant differences in vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) (Fig. 6, A and B). In addition, when compared with the HA-GEL transplantation group, the expression levels of proinflammatory cytokines [interferon- (IFN-)] were significantly decreased in the huMSCs/HA-GEL transplantation group (P < 0.01, Fig. 6C1), and significantly up-regulated expression was found for the anti-inflammatory cytokine [interleukin 4 (IL-4)] (P < 0.01, Fig. 6C2), as well as related cytokines that promote cell proliferation and tissue repair (IGF-1 and EGF) (P < 0.001, P < 0.05, respectively; Fig. 6, C3 and C4).

(A) The expression and localization of the potential cytokines secreted by huMSCs in the HA-GEL transplantation group and the huMSCs/HA-GEL transplantation group; inserted overview pictures show a lower magnification. (B) The optical density (OD) value of IGF-1, EGF, BDNF, VEGF, and HGF 2 months after HA-GEL transplantation or huMSCs/HA-GEL co-transplantation. (C1) Comparison of IFN- mRNA expression between the HA-GEL transplantation group and the huMSC/HA-GEL transplantation group. (C2) Comparison of IL-4 mRNA expression between the HA-GEL transplantation group and the huMSC/HA-GEL transplantation group. (C3) Comparison of IGF-1 mRNA expression between the HA-GEL transplantation group and the huMSC/HA-GEL transplantation group. (C4) Comparison of EGF mRNA expression between the HA-GEL transplantation group and the huMSC/HA-GEL transplantation group. *P < 0.05, **P < 0.01, and ***P < 0.001 versus 2 months postHA-GEL; the results shown are the mean SEM of three technical replicates from each animal.

At present, according to relevant reports, approximately 2.8 to 45.5% of women with impaired fertility suffer from IUA, and more than 90% of cases occur after pregnancy-related D&C (25). In this study, an IUA model was successfully established with an invasive surgery in nonhuman primates (rhesus monkeys), which have a genetic background, endocrine system, menstrual cycle, and anatomical structure that are similar to humans (26). This model allowed us to further explore new approaches for the intervention and treatment of adhesion, especially the thin endometrium caused by endometrial injury.

In the primate experiments, 6- to 7-year-old rhesus monkeys (reproductive age) were identified as the ideal subjects for establishing IUA models; they had regular menstrual cycles of approximately 21 to 30 days, which was observed and recorded in succession for the 2 months before mechanical injury. We developed the first model of endometrial injury in rhesus monkeys by open abdominal surgery; we verified the successful establishment by visualizing the hard and narrow cervix and confirmed that D&C could ultimately lead to severe IUA, which was characterized by severe endometrial fibrosis, loss of normal endometrial glands, paper-thin and discontinuous endometria, full of adhesive zone and endometrial cavity fluid, as well as amenorrhea, as noted by the American Fertility Society scoring method (27).

This study explored the effect of transplantation huMSCs combined with HA-GEL on intrauterine reconstruction and endometrial regeneration in an IUA model. In our study, when compared with the control group (transplantation of only HA-GEL), the effects of the group with huMSCs were confirmed by Doppler ultrasonic scanning, histological inspection, and ultrastructure detection 2 months after transplantation. In the experimental group, the injured endometrial tissue presented with a thicker endometrium, an increased number of endometrial glands, a decreased fibrotic area, and typical changes in the secretory phase, showing how the positive huMSCs acted upon endometrial repair and regeneration through secreting cytokines and growth factors (16); further, the HA-GEL acted as a physical barrier to severe adhesion and provided an ideal physical support for the attachment of huMSCs to prevent their rapid outflow from uterine cavity.

Specifically, in the control group (HA-GEL transplanted alone), subjects did not recover menstruation and normal uterine cavity. However, recovery of menstruation, the appearance of a normal uterine cavity, and normal cycling were observed in the other three rhesus monkeys 2 months after huMSCs/HA-GEL co-transplantation, suggesting the great effect of the huMSCs/HA-GEL complex on the reconstruction of the uterine cavity and on the blocking of adhesion. Meanwhile, we also found that HA-GEL transplantation could increase the number of endometrial glands, but it played no effective role in endometrial thickness, which was important because it was less effective than the huMSCs/HA-GEL complex. This amelioration of the damage to the endometria resulted in nearly normal levels and suggested the re-emergence of endometrial repair and regeneration after huMSCs/HA-GEL co-transplantation. In addition, the degree of fibrosis in the damaged endometria was found to be increasingly worse 2 months after transplantation despite HA-GEL or huMSCs/HA-GEL intervention, and it remained unclear whether HA-GEL had an effect on resisting fibrogenesis because of the small sample size of rhesus monkeys. Obviously, huMSCs/HA-GEL intervention relieved the worse aspects of fibrogenesis, suggesting a better outcome and potential effect on the reconstruction of abnormal tissue.

Then, it was unclear what the underlying mechanism of endometrial reconstruction was. Transplanted huMSCs were tracked in endometria, and the result showed no obvious labeled signal in endometrial tissue at 2 months after huMSCs/HA-GEL complex transplantation, which was contrary to a previous report (28, 29). To explain these conflicting results, three possibilities were proposed: (i) missed target area due to random sampling, (ii) completely eliminated following the outflow of menstrual blood, and (iii) huMSC apoptosis and depletion. However, the last two assumptions were preferred for the reason of multipoint sampling and continuous paraffin section, and if the endometrial tissues were obtained at 1 week or 2 weeks after huMSCs/HA transplantation, we might get different results owing to the similar menstrual cycles to human and different physiological function from animal models such as mice and rabbits. Alternatively, some cytokines and growth factors related to huMSCs were detected, and the results showed that huMSCs/HA-GEL complex transplantation could obviously increase the expression of IGF-1, EGF, BDNF, and so on compared to that of the control group (HA-GEL transplanted alone). Growth factors and their related peptides were deemed to mediate and regulate hormones working on target tissues through autocrine or paracrine function, and some growth factors, the endocrine basis of endometrium recycling including transforming growth factor, EGF, IGF, fibroblast growth factor, etc., were reported to regulate the differentiation and proliferation of endometrial cells (30). EGF, present in stromal and epithelial cells of the endometrium, could regulate endometrial proliferation, gland secretion, and decidual transformation (31). IGF played important roles in endometrial physiology and could regulate the cell cycle and promoted the proliferation of endometrial epithelial cells after the activation of estrogen (32, 33). Moreover, some reports showed the key role of BDNF in the regulation of endometrial cell proliferation by the downstream signal transducer and activator of transcription 3 signaling pathway and participating in the damage repair of endometrium (34, 35). Furthermore, considering the effect of huMSCs on uncontrolled fibrogenesis resulting from inflammatory activity and endometrial cell proliferation, anti-inflammatory cytokines (IL-4) were observed to be up-regulated, and proinflammatory cytokines (IFN-) were down-regulated; further, related cytokines that promote cell proliferation and tissue repair were up-regulated, such as IGF-1 and EGF, suggesting that excessive fiber formation could be inhibited by anti-inflammatory effects due to the advantageous microenvironment constructed by abundant huMSCs in the uterine cavity. VEGF, as the most important vascular growth factor, could be stimulated by ischemia and hypoxia in the endometrial layer after endometrial injury and played an important role in the early stage of endometrial repair and proliferation; during the time, angiogenesis could be promoted rapidly, but no effects were shown once the neovascularization was over (3639). We speculated that no difference found in VEGF expression might be related to the samples extracted from the endometrium during the secretory phase, a plateau stage of vascular repair and VEGF secretion in the endometrial basal layer. Overall, all of these results further verified the important role of huMSCs in damage repair by secreting a series of paracrine factors, such as anti-inflammatory factors, growth factors, and cytokines, related to constructing the microenvironment with properties such as anti-inflammatory, promoting repair, maintaining cell function, angiogenesis, etc., which was consistent with the previous report (40).

In conclusion, this study showed that both HA-GEL and huMSC/HA-GEL complexes could partially repair severe IUA caused by mechanical injury, but huMSC/HA-GEL complex transplantation indicated significant advantages in the dual repair effects of antiadhesive property and promotion of endometrial regeneration. By constructing a complex of huMSCs/HA-GEL with a biomaterial to prevent adhesion and allow stem cells to act at the appropriate site of repair of the endometrium, we have provided a method for solving a problem for patients with moderate to severe IUA and thin endometria caused by IUA. We hope that this novel strategy using the huMSCs/HA-GEL complex will be offered as a basic clinical research strategy in the future, and it might be a potential valuable treatment for gel loaded with cytokines to repair moderate to severe IUA.

The basic information for the six rhesus monkeys used in this study is shown in table S4. They were bred and supplied by Fujian Experiment Center of Nonhuman Primate for Family Planning, where they were maintained the clean-class animal feeding standards. We used these rhesus monkeys in the study following the outlined steps of the flowchart (fig. S4) from 17 September 2018 to 16 March 2019 with regular menstrual cycle observed (26). All experiments were conducted in accordance with the National Research Councils Guideline for the Care and Use of Laboratory Animals and approved by the Ethics Committee of the Center of Reproductive Medicine of Tongji Medical College of Huazhong University of Science and Technology in China.

The six rhesus monkeys were chosen for IUA models following mechanical injury, and intraperitoneal surgery was the proper choice for curettage in view of the special structure of the vagina and the cervix, which are characterized by hardness, toughness, a small aperture, and a long cervix. All monkeys were provided general anesthesia with an appropriate dose of ketamine by intramuscular injection, and then a mid-abdominal longitudinal incision was made to expose the uterus. A vertical incision (~0.5 cm) was made in the lower uterine segment. A small curettage spoon was used to deeply scrape the uterine walls until they became rough and pale, and the collected endometrial tissues were stored at 80C. Subsequently, the uterine and abdominal incisions were closed by continuous stitching with 6-0 Vicryl sutures and 3-0 silk absorbable sutures, respectively. After the operation, antibiotics were used to prevent infection, and observation on the first day after the operation showed that these rhesus monkeys were in good condition.

huMSCs were kindly provided by the Stem Cell Laboratory of the Center of Reproductive Medicine (Tongji Medical College, Huazhong University of Science and Technology, China). Frozen huMSCs between P3 and P9 were freshly seeded in 10-cm culture dishes (1 106 cells per dish) in Iscoves modified Dulbeccos medium (IMDM, Genom, China) supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, USA), penicillin (100 U/ml), and streptomycin (100 mg/ml; Gibco, USA). Briefly, the phenotypes of huMSCs were specifically identified by FACS, and the osteogenic and adipogenic capacities of the mesenchymal stem cells were assessed with a MesenCult Osteogenic Stimulatory Kit (STEMCELL Technologies Inc., Canada) and a MesenCult Adipogenic Differentiation Kit (STEMCELL Technologies Inc., Canada). P3 to P9 were used for the experiments.

To further evaluate the safety of huMSCs on HA-GEL, we mixed huMSCs (1 105 to 2 105 per well) and 300 l of HA-GEL [Con., 5 mg/ml; Bioregen, Co., Ltd., China, approved by CFDA as a medical device (no. 20153641542)] evenly with sterile syringes in 24-well plates and then added IMDM (Genom, China), including 10% (v/v) FBS (Gibco, USA), penicillin (100 U/ml), and streptomycin (100 mg/ml) (Gibco, USA). After co-culture for 48 hours, 0.8% collagenase type I supplemented with appropriate hyaluronidase was used to digest HA-GEL and release huMSCs, and FACS was chosen for the detection of cell apoptosis index in the coculture group (huMSCs/HA-GEL) and the culture-separated group (huMSCs), as well as live-dead cell detection with Live-Dead Cytotoxicity Assay Kit (MesGen Biotechnology, Shanghai).

Briefly, 50 l of huMSCs (1 107 to 2 107 cells) were injected into 200 l of HA-GEL (Bioregen, Co., Ltd. China), and then they were immediately transplanted into the uterine cavity through the open abdominal cavity. At the same time, the uterine cavity in the negative control group was injected into 200 l of HA-GEL following the same procedure that was used for the huMSC/HA-GEL transplantation group. All operations were performed under sterile conditions.

The endometrial thickness (before mechanical injury or after surgery for 2 months) was measured by an abdominal two-dimensional ultrasound system (Medison SA-600 Ultrasound System, Korea) with 3.5-MHz pulse repetition frequency to evaluate the damage to the endometrium and the endometrial regeneration.

The scraped pieces of endometrium were fixed in 4% paraformaldehyde for 24 hours and then embedded in paraffin. Serial paraffin-embedded sections (4 m) were obtained, sequentially dewaxed in xylene I and xylene II for 20 min each, and rehydrated in a series of ethanol solutions with a decreasing concentration (100% for 10 min, 100% for 10 min, 95% for 5 min, 90% for 5 min, 80% for 5 min, and 70% for 5 min). Then, the sections were rinsed in distilled water (three times, 5 min each). The sections were stained with an H&E solution (Servicebio, China) according to the manufacturers instructions. After staining, endometrial morphologic features were observed, and the number of uterine glands per unit area was counted according to five randomly selected high-power fields of each slide.

The 4-m paraffin sections of endometrium were dewaxed and rehydrated as described above and then were immersed in Masson A solution (Servicebio, China) overnight, which was followed by a brief wash under running water. Then, the sections were stained in a mixed solution of Masson A and Masson B (1:1, Servicebio, China) for 1 min, washed under running water, and placed in 1% hydrochloric acid alcohol for 10 s before they were washed again. Subsequently, sections were immersed in Masson D solution (Servicebio, China) for 6 min and then were stained in Masson E solution (Servicebio, China) for 1 min. The solution was then slightly drained, and the sections were placed directly in Masson F solution (Servicebio, China) for 2 to 30 s, and then they were rinsed in 1% glacial acetic acid for differentiation of the signals. Last, the sections were dehydrated in absolute ethyl alcohol, clarified in xylene for 5 min, and sealed in Permount Mounting Medium (Sinopharm Chemical Reagent Co., Ltd., China). Endometrial fibrosis was assessed according to five random fields on each slide, and the fibrotic area ratios were calculated using Image-Pro Plus software (version 6.0).

The total DNA samples were extracted from spleen tissue of rhesus monkeys and huMSCs carrying XY or XX chromosomes with a TIANamp Genomic DNA Kit (Tiangen Biotech Co., Ltd., Beijing). mRNA samples were extracted from endometrial tissue, and cDNA was synthesized with a RevertAid First Strand cDNA Synthesis Kit (Thermo, USA). Subsequently, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the specific expression of the DXZ1 gene on the X chromosome, SRY on the Y chromosome, and IFN-, IL-4, IGF-1, and EGF using StepOne and StepOnePlus Real-Time PCR Systems Version 2.3. The final reaction volume of 20 l contained 10 l of Bestar qPCR MasterMix (SYBR Green) (DBI Bioscience), 4 l of DNA samples, 0.4 l of forward/reverse primer (10 M), and 5.2 l of DNA/RNase-free double-distilled water (ddH2O). Last, agarose gel electrophoresis was performed to verify the expression of DXZ1 and SRY in the spleen and huMSC tissues. In addition, primer sequences used for DXZ1 (Xp11.1-Xq11.1), SRY (Yp11.3 Region), IFN-, IL-4, IGF-1, and EGF are summarized in table S5.

FISH analysis was performed to trace huMSCs for 2 months after the huMSCs/HA-GEL complexes were transplanted into the uterine cavity. Endometrial tissue was collected and immediately fixed in 10% paraformaldehyde before paraffin embedding. A Vysis SRY Probe LSI SRY Spectrum Orange/Vysis CEP X Spectrum Green Kit (Abbott Laboratories, USA) was used to mark huMSCs in the endometrium. All paraffin sections were dewaxed, rehydrated, hybridized with the probe, and so on according to the probes instructions; a final counterstaining of 4,6-diamidino-2-phenylindole (DAPI) was added, and visualization took place with fluorescence microscopy (Nikon Eclipse Ci, Nikon DS-U3). The orange (SRY/Y chromosome) and/or green (DXZ1/X chromosome) signals were used to verify the presence or absence of huMSCs in the endometrium.

The following factors were analyzed after transplantation of the huMSCs/HA-GEL complex: trophic factors (cytokines) that are secreted by huMSCs [according to relevant reports (17)], proinflammatory cytokines, anti-inflammatory cytokines, and related cytokines that promote cell proliferation and tissue repair. After fixation with 4% paraformaldehyde, paraffin embedding, and cutting 4-m paraffin sections, the slides were immersed in xylene and rehydrated through incubation in a series of alcohol gradients. The following specific antibodies were applied to sections at 4C overnight in humidified chambers: recombinant anti-BDNF antibody (EPR1292) (ab108319, Abcam), anti-VEGF antibody (C-1: sc-7269, Santa Cruz), antiIGF-1 antibody (W18: sc-74116, Santa Cruz), anti-HGFa antibody (H-10: sc-374422, Santa Cruz), and anti-EGF antibody (F-9: sc-166779, Santa Cruz). Then, these sections were incubated with a Cy3-tagged secondary antibody for 1 hour at room temperature and were then rinsed in ddH2O three times. Counterstaining was performed with DAPI for 5 min, and the fluorescence signal was detected under a fluorescence microscope (Nikon Eclipse Ci, Nikon DS-U3).

We collected three technical replicates from each animal and repeated the experiments at least three times. The data presented as the mean SEM were analyzed with Statistical Package for the Social Sciences Statistics 17.0. The normally distributed numerical variance was assessed by a t test with homogeneity of variance, and 2 tests were used to analyze the differences between two or more rates. The percentage of positive area after H&E staining and Masson staining was measured using ImageJ 1.43u (Wayne Rasband, National Institutes of Health, USA). Statistical significance was assumed for P < 0.05.

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

Acknowledgments: We acknowledge the breeding and technical assistance of Fujian Experiment Center of Nonhuman Primate for Family Planning. We thank C. Yu for the operation of uterine D&C and open abdominal surgery. We thank C. Xiong and P. Su for their invaluable contributions in critically revising the manuscript and providing guidance for important intellectual content. We thank M. Zhang, T. Chang, and S. Song for analyzing the data and collecting the samples. Funding: This work was supported by the National Natural Science Foundation of China (NSFC 81571434) and the National Key Research and Development Program of China (2017YFC1002002). The funders played no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Author contributions: W.X. designed the study and published this manuscript; L.W. and C.Y. performed the experiments and wrote the manuscript; M.Z., T.C., and S.S. analyzed the data and collected the samples; C.X. and P.S. provided their invaluable contributions in critically revising the manuscript and providing guidance for important intellectual content. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

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In situ repair abilities of human umbilical cordderived mesenchymal stem cells and autocrosslinked hyaluronic acid gel complex in rhesus monkeys with...

UH studying use of convalescent plasma to improve health of COVID-19 patients – Crain’s Cleveland Business

University Hospitals is participating in a study to determine if plasma donated from someone who's recovered from COVID-19 can improve the health of patients battling the virus.

Because patients who've recovered from COVID-19 have antibodies in their blood that could help fight the virus, it is thought that those suffering complications from COVID-19 might improve faster if they receive plasma from the people who've recovered, according to a news release.

Headed at the Christ Hospital in Cincinnati, the study currently has three participating sites in Ohio. Being a part of the study enhances UH's ability to quickly get plasma for patients who need it, according to the release.

"UH is at the forefront of experimental treatments including remdesivir, stem cell therapy and now convalescent plasma," cardiologist Dr. Steven Filby said in a prepared statement. "Patients suffering from extreme complications of COVID-19 have hope at UH thanks to these options."

Filby is a co-investigator for the study, along with Dr. Eiran Gorodeski, an internist and cardiologist at UH Cleveland Medical Center, and Dr. Katharine Downes, a pathologist.

Symptoms of COVID-19 include fever, fatigue, dry cough, headache and more. In addition to respiratory distress, COVID-19 can affect the cardiovascular system. No FDA-approved medicine currently exists to treat or prevent COVID-19, according to the release.

To receive plasmas as part of this study, patients must be hospitalized with COVID-19 and be experiencing "serious complications," including myocardial injury, according to the release.

Blood donations will be collected from patients who are determined recovered and cleared from COVID-19 and who are found to have developed antibodies in their plasma. UH is partnering with Hoxworth Blood Center and Vitalant to provide donations for the study.

"Offering experimental COVID-19 convalescent plasma transfusion to our patients gives us another option to fight COVID-19," Downes said in a prepared statement. "UH appreciates our partnership with the blood suppliers that are crucial to making this happen."

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COVID-19: Recommendations for Treating MS and Related… : Neurology Today – LWW Journals

Article In Brief

Most therapies for multiple sclerosis and neuromyelitis optica spectrum disorder should be continued and not stopped during the COVID-19 pandemic, MS experts agree. But there are also pros and cons of starting certain B-cell depleting and other therapies.

For most patients with multiple sclerosis (MS), the benefits of staying on therapy will outweigh the risks of stopping due to concerns over COVID-19, according to new guidelines for treating MS during the pandemic.

Even in those with a documented mild case of COVID-19, continued treatment with most MS medications may be reasonable, the guidelines recommend. However, they emphasize: Neurologists should have a lower threshold for stopping treatment in people taking therapies with greater immunosuppressive effects and those with risk factors for a more severe disease (older age, comorbidities), or if COVID-19 symptoms are deteriorating.

Published online in Neurology on April 2 by a team of MS neurologists from the United States, Australia, The Netherlands, and the United Kingdom, the new guidelines cover both MS and neuromyelitis optica (NMO) spectrum disorder. The guidelines differ only slightly from guidelines previously released by the National Multiple Sclerosis Society, the Italian Society of Neurology, and other groups.

Even so, MS neurologists not involved in preparing the recommendations welcomed their publication.

Kudos to the authors for taking the time to do this when we're all under so much pressure, said Annette Langer-Gould, MD, PhD, the regional lead for clinical and translational neuroscience for the Southern California Permanente Medical Group/Kaiser Permanente.

Based on prior experience with other viral infections in people with MS, the recommendations will likely need to be updated as data emerges from actual cases of patients who develop COVID-19.

New data are emerging quickly from clinical experience and from registries that have been established for MS patients with COVID-19, said the first author of the paper, Wallace Brownlee, MD, PhD, a neurologist with the Queen Square MS Centre and the National Hospital for Neurology and Neurosurgery in London.

Indeed, one recommendation in the paper is already out of date. Face masks are only recommended for people who are coughing or sneezing, or for those caring for a patient with suspected COVID-19 infection, the guidelines stated. By now, of course, most public health recommendations call for wearing face masks whenever people are in public and are unable to stay at least six feet away from others.

Otherwise, MS neurologists told Neurology Today that they had few if any disagreements with the guidelines as published. In particular, they all agreed that IV treatments with drugs known to cause significant declines in immune function should be avoided or delayed as long as possible.

Dr. Brownlee and other MS neurologists urged any neurologist treating a patient with MS who develops a COVID-19 infection to submit data to one of the patient registries that have been established. In North America, the National MS Society and the Consortium of MS Centers have established the Coronavirus and MS Reporting Database at http://www.covims.org.

For MS patients who are just beginning treatment, Dr. Brownlee said, We recommend that neurologists take a cautious approach to initiating patients on treatments that can be associated with periods of significant immune suppression, including autologous hematopoietic stem cell transplantation, alemtuzumab and cladribine.

Although acute MS relapses are often treated with a short course of high-dose IV methylprednisolone, such treatments should be avoided during the pandemic, the guidelines stated. High-dose steroids hasten the recovery from MS relapses, but do not influence the final degree of recovery, the paper noted. Because steroids can increase the risk of infection, neurologists should have a higher threshold for offering them during the COVID-19 pandemic, according to the guidelines.

A few disease-modifying therapies (DMTs), including interferon-beta and glatiramer acetate, do not increase the risk of systemic infections. Other DMTs, however, do have immunosuppressive effects with alterations in lymphocyte number, trafficking, proliferation and function, with an increased risk of infections, including viral infections and respiratory infections, the guidelines stated.

People with MS who are profoundly lymphopenic, for example, after treatment with alemtuzumab or less commonly during treatment with cladribine, fingolimod or dimethyl fumarate, may be at higher risk.

As reasonable as such concerns appear to be at this time, the paper noted that no data specific to MS patients with COVID-19 has yet emerged supporting them.

For patients scheduled for routine treatment with alemtuzumab or cladribine, We recommend delaying treatment with these therapies, the paper stated. Likewise, standard every six-month dosing with ocrelizumab or rituximab can also be delayed in most cases.

B-cell depletion frequently lasts much longer than the scheduled dosing interval, the recommendations noted. Extended interval dosing should be considered, especially in patients who are B-cell depleted...or [in] those with low levels of immunoglobulin-G. Extended interval dosing is already widely used in patients treated with natalizumab because of observational data showing a reduced risk of progressive multifocal leukoencephalopathy. Whether this approach reduces the risk of other infections is unknown but should be considered during the COVID-19 pandemic to reduce hospital visits.

For MS patients who are hospitalized with a severe COVID-19 infection, consideration should be given to stopping treatment, the guidelines state. Treatment can be restarted after four weeks, or when symptoms have fully resolved, keeping in mind the risk of rebound MS activity with S1P modulators and natalizumab. Neurologists should alert intensive care physicians to the importance of fever management in people with MS.

Patients with neuromyelitis optica spectrum disorder who do not have a COVID-19 infection should be encouraged to continue attack-prevention therapies, because relapses of NMOSD can be devastating. If the need to stop or delay treatment in such patients arises, then moderate dose corticosteroids (e.g. prednisolone 20mg) can be used to prevent relapses in the short to medium term, the guidelines recommended.

Dr. Langer-Gould echoed the guidelines' concern about MS drugs associated with lymphopenia.

With COVID-19, we're seeing something very unusualthat in the people who do poorly, almost all of them have lymphopenia when they're admitted, she said. Any drug you're on that is causing T-cell lymphopenia is more likely to increase your risk of getting a severe case of COVID. So they have correctly identified the ones to stay away from, including alemtuzumab and cladribine, but I would add fingolimod, dimethyl fumarate and other S1P modulators to that list.

Starting in early March, she said, We actively reached out to patients on dimethyl fumarate and S1P inhibitors and are switching them, depending on their disease severity, either to interferon and glatiramer acetate, or if they had active disease, then we switched them to either rituximab or natalizumab.

But for all her patients other than those on interferon-beta or glatiramer acetate, she said, We're telling them to consider themselves immune-suppressed. They should immediately get themselves tested for COVID-19 if they develop fever or shortness of breath.

Her practice recently had an MS patient who had been stable on natalizumab until suddenly developing a high titer positive antibodies against JC Virus. The patient is now at increased risk of progressive multifocal leukoencephalopathy, Dr. Langer-Gould said.

Our plan had been to switch them to rituximab, but then they developed a COVID-19 infection and are mildly symptomatic. That's a big problem, because you need to pre-treat for rituximab with steroids, potentially increasing the risk of a more severe case of COVID-19, and the infusion is long, which raises the risk of infecting the nursing staff at the infusion center. We've decided to give her another dose of natalizumab eight weeks after her last dose. It's a short infusion, and we don't have to pre-treat with steroids. But what's the right decision?

Timothy L. Vollmer, MD, FAAN, professor of neurology at the University of Colorado Health Sciences Center and medical director of the Rocky Mountain MS Center, said that most of his group's patients are on ocrelizumab.

We probably didn't need to be dosing patients every six months, he said. As a result of COVID-19, we're reevaluating the dosing strategy. We're checking their B cells and antibody levels, and if they are still depleted, we wait another two months. It will take some patients a year or more before they begin to normalize their B cells. Dosing less frequently will also decrease costs substantially and make the drug more attractive for patients to use.

One clear effect of the COVID-19 pandemic is that many MS patients are reaching out to their neurologists about what it means for them.

My colleagues and I have been receiving many phone calls and messages about our recommendations, said Bardia Nourbakhsh, MD, assistant professor of neurology at Johns Hopkins. We try to extrapolate the information that we have from other viral infections.

Dr. Nourbakhsh said he had been contacted the week of April 13 by a community neurologist whose patient had developed a severe, disabling, demyelinating relapse that did not improve after IV steroids.

My recommendation was for the neurologist to set up a plasma exchange, he said. I would not delay the treatment of a serious relapse that could affect the mobility or vision of the patient. Treating an actual condition takes precedence over the possibility of coming into contact with coronavirus.

Bianca Weinstock-Guttman, MD, director of the Jacobs MC Center at the University of Buffalo and director of its Pediatric MS Center of Excellence, said she recommends that all MS patients should be reminded, now more than ever, to follow basic daily health recommendations.

Supportive recommendations for patients include keeping interactions with friends and relatives through video, exercising via YouTube, maintaining a healthy diet, and vitamin supplementation, especially vitamin D, she said.

Joseph Berger, MD, FAAN, professor and associate chief of the Multiple Sclerosis Division at the University of Pennsylvania School of Medicine, noted that the very same tendency of some MS drugs to suppress immune function could in fact benefit COVID-19 patients who develop acute respiratory distress syndrome (ARDS).

Individuals who end up developing ARDS are not dying because of unsuppressed viral replication, Dr. Berger said. Rather, it's an overly robust immune response, a cytokine storm, that appears to give rise to the ARDS. Many of the drugs we use may actually have a beneficial effect on the phase of the illness that results in high morbidity and mortality.

He pointed out that a non-randomized, open-label trial is underway in China to test the effect of giving fingolimod to 30 patients with COVID-19 in order to prevent ARDS.

Dr. Weinstock-Guttman said another drug that might block the cytokine storm is tocilizumab, approved to treat moderate to severe rheumatoid arthritis.

IL-6 blockade was shown beneficial recently also for NMO patients, so it will be interesting to see COVID-19 patients' outcome when treated with antiIL-6 products for a previous underlying disease, Dr. Weinstock-Guttman said.

Dr. Langer-Gould expressed caution about the approach. There's a big difference between using a drug like fingolimod for a few days to reduce a cytokine storm versus having someone on it as a long-term therapy, which results in chronic T-cell suppression and, in rare instances, fatal viral infections, she said.

If your T-cells are markedly diminished and you are infected with COVID-19, your body would have a hard time clearing that virus and you would potentially be at higher risk of developing pneumonia, ARDS, and, potentially, multisystem organ failure.

She added that none of the immunologists she works with are recommending a T-cell-depleting therapy for COVID-19. Most of the patients we're seeing already have low lymphocytes, she said. The virus is taking down the CD4 and CD8 cells. I don't think anyone would feel comfortable giving a drug that further suppresses CD4 and CD8.

Dr. Brownlee agreed that while the potential benefit of fingolimod as an acute treatment to prevent ARDS is interesting, we need to be careful about being too quick to translate hypotheses into treatment. It's not enough to inform patient care at the moment.

Ultimately, such questions can only be answered as more experience is gained in treating MS patients who develop a COVID-19 infection, Dr. Berger said. Time will tell, he said. It's going to be important to get real-world data from the registries to see whether or not what we think is correct. Is there a signal for any of these MS drugs? We'll know when the registries reveal their data. It's going to take the participation of neurologists around the world to distill out the treatments with one or another drug.

Dr. Brownlee has accepted speaker honoraria and/or participated in advisory boards for Biogen, Merck, Mylan, Novartis, Roche and Sanofi-Genzyme. Dr. Vollmer has received compensation for lectures and consultancy with Biogen IDEC, Genentech/Roche, Siranax, Celgene, EMD Serono, and Novartis. He has received research support from Rocky Mountain MS Center, Biogen,Actelion, Roche/Genentech;, F. Hoffman-La Roche, Ltd., and TG Therapeutics, Inc. Dr. Berger has received honoraria and an institutional grant from Biogen, and Genentech/Roche. He has received honoraria as a consultant for Celegene, Millennium/Takeda, Novartis, Inhibikase, Excision Biom Amgen, Shire, Dr. Reddy, Serono, Morphic, Encycle, Merck, and MAPI. Dr. Nourbarkhsh served on the advisory board for Jazz Pharmaceutical. Dr. Langer-Gould had no disclosures.

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New hybrid embryos are the most thorough mixing of humans and mice yet – Science News

Scientists have made embryosthat are a lot mouse and a little bit human.

With a little help, human stem cells can knit themselves into growingmouse embryos, populating thedeveloping liver, heart, retina and blood, researchers report May 13 in Science Advances.

Finicky human cells dont tend to grow well in other animals. But in one of the new mouse embryos, 4 percent of its cells were human the most thorough mixing between human and mouse yet.

That level of integration isquite striking to me, says Juan Carlos Izpisua Belmonte, a stem cell anddevelopmental biologist at the Salk Institute for Biological Studies in LaJolla, Calif. If other scientists can replicate the findings, it potentiallyrepresents a major advance, says Izpisua Belmonte, who was not involved in thestudy.

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Such chimeras could helpreveal how a single cell can give rise to an entire organism. More humanizedanimals could also prove valuable in studying diseases such as malaria that affectpeople more than other animals. And with more advances, chimeras couldultimately turn out to be a source of human organs.

Many scientists have hitroadblocks in growing human stem cells in mice or other animals, including pigs and cows(SN: 1/26/17). We have analyzedthousands of embryos but never saw robust chimeric contribution of human stemcells to mouse embryos beyond day 12, says stem cell and developmentalbiologist Jun Wu of the University of Texas Southwestern Medical Center inDallas, who wasnt involved in the study.

The new methods success comes down to timing, says neuroscientist and stem cell biologist Jian Feng. To grow and thrive in a mouse embryo, human stem cells developmental clocks must be turned back to an earlier phase called the nave stage. You need to basically push the human cells back to that phase, says Feng, of the University at Buffalo in New York.

Feng and his colleagues resetthe stem cells clocks by silencing a protein called mTOR for three hours. Thisbrief treatment shocked the cells back to their nave stage, presumably restoringtheir ability to turn into any cell in the body.

Researchers injected batchesof 10 to 12 of these more youthful human stem cells into mouse embryos containingabout 60 to 80 mouse cells, and allowed the embryos to develop for 17 days.

To outward appearances, these embryos grew normally despite harboring human cells. By tallying DNA that was specific to either mouse or human, the researchers found that human cells accounted for between 0.1 and 4 percent of the total cells in the embryos.

Human cells knittedthemselves into most developing tissues of the mouse, destined to become theliver, heart, bone marrow and blood. Human red blood cells were particularlyabundant in these mouse embryos, the researchers found. A small number of humancells showed up in tissue that will form a brain; one embryo had a swarm of humanphotoreceptors, eye cells that help detect light.

As far as the researcherscould tell, no human cells were among the cells that go on to form sperm andegg. The capacity of chimeras to reproduce is one of the worrisome ethicalquestions surrounding the organisms that scientists are still trying to figureout.

Once inside a mouse embryo, the normally sluggish developmental pace of the human cells sped up to match their hosts. Human stem cells typically are slow to turn into certain types of mature photoreceptors, liver cells or red blood cells, Feng says, but not when the human cells are inside a mouse embryo. You put the same human cells in a mouse embryo, [and] they go fast, Feng says. In 17 days, you get all these mature cells that would otherwise take months to get in a normal human embryo.

Other scientists emphasize that different laboratories need to repeat the results. But if it works a big if here this has big implications, Wu says.

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Hotel in Spain requires two negative Covid-19 tests to check in with a free antibody test during your stay – CNBC

Travelers the world over are asking: Where can I travel that's safe?

Sha Wellness Clinic, a medical clinic and hotel in southeastern Spain, has an answer. It's requiring guests to provide a negative Covid-19 test result before they ever step foot in the door.

"When we began to put together our preventative measures, there was no doubt that this would be a requirement," said Alejandro Bataller, vice president of the clinic. "Our main priority is ensuring our guests' safety while at Sha, so they can have peace of mind during their stay."

Covid-19 and antibody tests are free of charge to guests at Sha Wellness Clinic.

Courtesy of Sha Wellness Clinic

The medicalclinic, which is located in Alicante, Spain, is banking on the fact that requiring guests to get a coronavirus test before they arrive is the very thing that will make guests want to come.

"During these times, people are more concerned than ever about their safety when it comes to traveling," Bataller said. "Guests can have some peace of mind knowing that everyone visiting has been tested, and the spaces are constantly being sanitized, so they can truly relax and feel confident and safe."

All guests must undergo Covid-19 testing 24 to 48 hours before arriving at Sha Wellness Clinic. Guests are encouraged to send the results, though Sha will allow them to bring the results when they arrive too.

Guests are not the only ones being tested. The clinic staff is required to get tested for Covid-19 before returning to work and regularly thereafter.

Upon arrival, all guests will undergo a second Covid-19 test as well as an antibody test and a medical examination. All testing is included with a stay free of charge.

"It is important to us that our guests not only feel safe when returning to Sha, but also feel healthier during their stay," said Bataller.

In line with post-pandemic changes happening across the hotel industry, the clinic is intensifying its cleaning protocol, installing thermographic cameras to detect body temperatures and eliminating all paper touch points, such as food and spa menus.

Sha Wellness Clinic is located in Alicante, Spain.

Courtesy of Sha Wellness Clinic

UV disinfection towers will be installed in the clinic suites, and ozone treatments will be used to disinfect vehicles and luggage. The check-in process is being moved into guest suites.

As part of all stays, guests embark on health programs that focus on topics like fitness, detoxing and weight loss. The clinic has 11 residences where guests can undergo treatments and medical consultations without having to leave their suites.

The clinic is reducing occupancy levels in common areas to encourage social distancing.

Courtesy of Sha Wellness Clinic

Sha is adding to its immune system treatments in response to the pandemic and adding new "immune system strengthening" services. Guests can undergo stem cell therapy, infrared heat, vitamin C megadoses, thermal shock reinforcement and stress management sessions, to name a few. Immunotherapy and lymphocyte profile consultations are now included in all bookings as well.

A seven-day booking for the "rebalance" program in late July in the entry-level deluxe suite is approximately 5,310 euros (US$5,745). To upgrade to a bi-level garden residence, the rate jumps to 17,000 euros (US$18,390) and nearly twice that figure for a stay in the penthouse residence.

The Waldhotel, a medical wellness resort in Obbrgen, Switzerland, is testing guests for Covid-19, though it does not require a negative test result before guests can check in.

Tests cost 150 Swiss francs (US$154) and are administered in the Waldhotel Health & Medical Excellence, a medical center that operates in the hotel.

"Although the Waldhotel Health & Medical Excellence remained open throughout the pandemic, no Covid-19 case has been registered in the Waldhotel and the Brgenstock Resort," said Dr. Verena Briner, the medical director of the center, referencing the larger resort in which the Waldhotel resides.

The Waldhotel is part of the Brgenstock Hotels & Resort, near Switzerland's Lake Lucerne.

Courtesy of Waldhotel

The Waldhotel has Covid-19 antibody testing for guests for 100 Swiss francs (US$102) with results returned in one day.

As the Waldhotel is a five-star hotel and medical center in one, the hotel says it had strict hygiene measures in place such as hand disinfection stations and housekeeping protocols before the pandemic started. Still, it instituted new measures in response to the virus under the guidance of Dr. Briner.

The Waldhotel Health & Medical Excellence center operates inside the Waldhotel.

Courtesy of Waldhotel

The medical center employs a team of experts in dermatology, gastroenterology, sports medicine and dentistry. Hotel guests can join guided programs that focus on exercise, aesthetics, mindfulness, immunity and weight loss. Programs range from three days to two weeks, though some guests stay for months.

The hotel has access to an airfield where private jets can land for guests who can afford to avoid commercial airlines.

International travel to Spain and Switzerland isn't possible right now, but it may soon be, depending on where you live.

After two of months of strict lockdown, Spain began to lift nationwide stay-at-home guidelines in May. Currently, Spain has a 14-day quarantine requirement for most people arriving from abroad, though Spain's Transport Minister Jose Luis Abalos said he hopes tourism activity will start again in late June.

The lobby of the Waldhotel.

Courtesy of Waldhotel

Sha Wellness Clinic closed at the end of March and is aiming to reopen in July, pursuant to guidelines set forth by the Spanish government.

Right now, only Swiss citizens, residents and select workers can enter Switzerland. However, Switzerland announced intentions to broadly reopen its borders with Germany, Austria and France on June 15. There are no plans to reopen borders with Italy yet.

On May 14, Swiss International Air Lines announced it would restart flying up to 190 flights from Zurich and Geneva in June. Flights will resume in stages and will go to 41 European destinations including Paris, Rome, Barcelona and Copenhagen. Additionally, the airline is providing new intercontinental direct connections in June, linking Zurich to New York, Singapore, Tokyo and Johannesburg, among other cities.

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‘Outpatient Is Still Chaos’: What We Heard This Week – MedPage Today

"We still predominately test if you are being admitted or, now, if you need an elective surgery or admission to a skilled nursing facility. Outpatient is still chaos." -- An anonymous infectious diseases specialist, responding to a MedPage Today benchmark survey about challenges healthcare workers face during the COVID-19 pandemic.

"They are a harbinger of what is to come. In several decades, these conditions will start affecting large regions and for longer periods of time." -- Matthew Huber, PhD, of Purdue University in West Lafayette, Indiana, about potentially fatal heat and humidity mixtures recorded at weather stations throughout the world.

"Don't buy any fancy cars." -- Andrew Freedman, MD, program director of the urology residency program at Cedars-Sinai Medical Center in Los Angeles, on his advice to medical trainees in the current uncertain economic climate.

"It's actually incredibly complicated." -- Alysse Wurcel, MD, of Tufts Medical Center in Boston, on the logistics of testing prison and jail inmates for coronavirus infection.

"It is an important milestone in the field as it reports on survival of stem cell-derived dopamine neurons in a human brain." -- Malin Parmar, PhD, of Lund University in Sweden, discussing a novel brain cell transplant performed in a Parkinson's disease patient.

"Children are coming in critically ill, with fever and abdominal pain, vomiting and in shock." -- Nadine Choueiter, MD, of the Children's Hospital at Montefiore, on the multi-system inflammatory disorder in children apparently connected to COVID-19.

"It definitely is taking a lot of patients a decent amount of time to feel that they are perfectly back to normal." -- Aaron Glatt, MD, chair of the department of medicine at Mount Sinai South Nassau, on recovery from COVID-19.

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Sumit Dewanjee, MD, is being recognized by Continental Who’s Who – PRNewswire

TEMPE, Ariz., May 7, 2020 /PRNewswire/ --Sumit Dewanjee, MD, is being recognized by Continental Who's Who as a Top Orthopedic Surgeon in the field of medicine as an at FXRX Orthopedics and Bracing.

Serving patients in the metropolitan Phoenix, Arizona area FXRX is the leading practice in Arizona for orthopedic surgery. FXRX mainly focuses on orthopedic sports injuries of the hip, knee, and shoulder, along with orthopedic fracture treatments such as stem cell therapy and cartilage restoration procedures.

Dr. Sumit Dewanjee is the founder and premier Board-certified Orthopedic Surgeon at FXRX with 16 years' experience in orthopedic surgery. His particular areas of focus are knee and shoulder surgery, fracture treatment and he's knowledgeable in both general orthopedics and orthopedic bracing. His additional skills include performing minimally invasive procedures using arthroscopy, with expertise in minimizing blood loss and patient recovery periods while providing the results and benefits of advanced and cutting edge treatments in surgery.

To gain the knowledge needed for his successful career with his private practice, Dr. Dewanjee attended the University of Miami and received his Medical Degree followed by Maricopa Medical Center to complete his residency.

In addition to his affiliations with MENSA, American Academy Ortho Surgeons, Dr. Dewanjee has been awarded the Patient Choice Award for the 7th time, maintaining his status as one of the best orthopedic surgeons in Phoenix, AZ. and Compassionate Doctors.

During his free time, Dr. Dewanjee donates to The Human Society.

Dr. Dewanjee dedicates his success to his mentor Dr. Dana Seltzer.

For more information please visit fxrxinc.com

Contact: Katherine Green , 516-825-5634 [emailprotected]

SOURCE Continental Who's Who

http://www.continentalwhoswho.com

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Results From Pivotal Phase 2/3 Study of Emapalumab in Patients With Primary HLH Published in New England Journal of Medicine – BioSpace

STOCKHOLM, May 7, 2020 /PRNewswire/ -- Sobi announced today that the results from the pivotal phase 2/3 study evaluating the efficacy and safety of emapalumab in patients with primary haemophagocytic lymphohistiocytosis (HLH) were published in the New England Journal of Medicine on 7 May 2020. Emapalumab is the first therapy approved by the US Food & Drug Administration (FDA) for primary HLH and is under review by the European Medicines Agency (EMA).

Primary HLH is a rare syndrome that typically presents in infancy but can also be seen in adults and is associated with high morbidity and mortality. This life-threatening disease is characterised by immune dysregulation and uncontrolled hyperinflammation. The treatment objective is to suppress the hyperinflammation and control the acute features of the disease in order to successfully bring patients to haematopoietic stem cell transplantation (HSCT).

"The publication of the results in this highly respected medical journal is a testament to the medical importance of the emapalumab findings for patients with primary HLH," says Milan Zdravkovic, Head of Research & Development and Chief Medical Officer at Sobi. "The results further advance our understanding of primary HLH and the role of interferon gamma in its pathogenesis. Our hope is to contribute to the improvement of care and treatment for patients suffering from this potentially fatal disease."

The results with emapalumab in primary HLH published in the New England Journal of Medicine highlight the overall response rate of 63 percent in previously treated patients at the end of up to 8 weeks of treatment (compared to the pre-specified null hypothesis of 40 percent (p=0.02)). In the previously treated group, 70 percent of patients were able to proceed to transplantation. The most commonly reported adverse reactions ( 20 per cent) were infections, hypertension, infusion-related reactions and fever.

Michael Jordan, Professor of Pediatrics at the Cincinnati Children's Hospital Medical Center in the US and coordinating Principal Investigator of the study (US), confirms the importance of making advances in finding new therapies for HLH and emphasises the importance of the publication of the results: "The findings from the study are encouraging for those affected by this devastating disease."

Professor Franco Locatelli, Head of the Department of Onco-Haematology, Bambino Ges Children's Hospital IRCCS, Sapienza University of Rome, Italy, and coordinating Principal Investigator (EU), adds: "Emapalumab represents a prototype model molecularly targeted therapy and an important step towards improving outcomes for this severe and life-threatening disease."

This pivotal clinical study is the first study in primary HLH to prospectively assess and report treatment responses using predefined comprehensive objective clinical and laboratory criteria. Preclinical data have shown the central role of interferon gamma (IFN) in the pathogenesis of this disease1. Emapalumab is a monoclonal antibody that binds to and neutralises IFN. It was approved by the US Food & Drug Administration (FDA) on the basis of this clinical study for the treatment of primary HLH in adult and paediatric (newborn and older) patients with refractory, recurrent or progressive disease, or intolerance to conventional HLH therapy, and received Breakthrough Designation prior to review.

About emapalumab

Emapalumab is a monoclonal antibody that binds to and neutralises interferon gamma (IFN). In the US, emapalumab is indicated for paediatric (newborn and older) and adult primary haemophagocytic lymphohistiocytosis (HLH) patients with refractory, recurrent or progressive disease, or intolerance to conventional HLH therapy. Emapalumab is the first and only medicine approved in the US for primary HLH, a rare syndrome of hyperinflammation that usually occurs within the first year of life and can rapidly become fatal unless diagnosed and treated. The FDA approval is based on data from the phase 2/3 studies (NCT01818492 and NCT02069899). Emapalumab is indicated for administration through intravenous infusion over one hour twice per week until haematopoietic stem cell transplantation (HSCT). For more information please see http://www.gamifant.com including the full US Prescribing Information. Emapalumab is under review for primary HLH by the European Medicines Agency (EMA).

About SobiTM

Sobi is a specialised international biopharmaceutical company transforming the lives of people with rare diseases. Sobi is providing sustainable access to innovative therapies in the areas of haematology, immunology and specialty indications. Today, Sobi employs approximately 1,400 people across Europe, North America, the Middle East, Russia and North Africa. In 2019, Sobi's revenues amounted to SEK 14.2 billion. Sobi's share (STO:SOBI) is listed on Nasdaq Stockholm. You can find more information about Sobi at sobi.com.

For more information please contactPaula Treutiger, Head of Communication & Investor Relations+ 46-733-666-599paula.treutiger@sobi.com

Linda Holmstrm,Corporate Communication & Investor Relations+46-708-734-095linda.holmstrom@sobi.com

1. Jordan et al. Blood 2004;104:735-43.

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SOURCE Swedish Orphan Biovitrum AB

Company Codes: Berlin:B6E, Bloomberg:SOBI@SS, ISIN:SE0000872095, LSE:0MTD, OTC-PINK:BIOVF, RICS:SOBI.ST, Stockholm:SOBI, Dusseldorf:B6E, Frankfurt:B6E, Stuttgart:B6E

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Results From Pivotal Phase 2/3 Study of Emapalumab in Patients With Primary HLH Published in New England Journal of Medicine - BioSpace

COVID-19 Impact and Recovery Analysis | Global Cell Isolation Market 2020-2024 | Evolving Opportunities with Agilent Technologies Inc. and Becton,…

LONDON--(BUSINESS WIRE)--The global cell isolation market size is expected to grow by USD 9.47 billion as per Technavio. This marks a significant market growth compared to the 2019 growth estimates due to the impact of the COVID-19 pandemic in the first half of 2020. Moreover, steady growth is expected to continue throughout the forecast period, and the market is expected to grow at a CAGR of almost 21%.

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With the spread of COVID-19, a global increase in R&D activities in AR and PB has been observed in the first half of 2020. Researches are being conducted on stem cell therapies to develop mesenchymal stem cell-based therapies. For instance, in March 2020, researchers from the University of North Texas Health Science Center initiated a clinical trial in China on more than 100 COVID-19 patients and administered injections of mesenchymal stem cells. The rise in such R&D activities is increasing the demand for cell isolation products, which will result in the growth of the cell isolation market during the forecast period.

Technavio's custom research reports offer detailed insights on the impact of COVID-19 at an industry level, a regional level, and subsequent supply chain operations. This customized report will also help clients keep up with new product launches in direct & indirect COVID-19 related markets, upcoming vaccines and pipeline analysis, and significant developments in vendor operations and government regulations.

https://www.technavio.com/report/global-cell-isolation-market-industry-analysis

The market is driven by the increasing prevalence of chronic diseases. In addition, product advances are anticipated to boost the growth of the cell isolation market.

With the growing incidence of chronic diseases globally, the applications of isolated human cells in research, drug development, and clinical trials are increasing. Cell isolation helps study the behavior of cells and their response to diseases and different drugs. Cell isolation tools are used in drug discovery to generate recombinant protein therapeutics, which can treat a variety of chronic diseases. Hence, the rising prevalence of chronic diseases will boost the need for cell-based therapies, which, in turn, will increase the demand for cell isolation tools.

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Major Five Cell Isolation Companies:

Agilent Technologies Inc.

Agilent Technologies Inc. has business operations under various segments, such as life sciences and applied markets, diagnostics and genomics, and Agilient CrossLab. The company offers various consumables and instruments for cell isolation procedures.

Becton, Dickinson and Co.

Becton, Dickinson and Co. operates its business through various segments, such as BD Medical, BD Life Sciences, and BD Interventional. The company provides consumables and instruments for cell isolation procedures.

Bio-Rad Laboratories Inc.

Bio-Rad Laboratories Inc. has business operations under two segments: life science and clinical diagnostics. The company provides consumables and instruments for cell isolation procedures.

Danaher Corp.

Danaher Corp. operates its business through various segments, such as life sciences, diagnostics, and environment & applied solutions. The company provides consumables and instruments for cell isolation procedures.

F. Hoffmann-La Roche Ltd.

F. Hoffmann-La Roche Ltd. has business operations under two segments: pharmaceuticals and diagnostics. The company provides consumables and instruments for cell isolation procedures.

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Cell Isolation Market Product Outlook (Revenue, USD bn, 2020-2024)

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With over 500 specialized analysts, Technavios report library consists of more than 17,000 reports and counting, covering 800 technologies, spanning across 50 countries. Their client base consists of enterprises of all sizes, including more than 100 Fortune 500 companies. This growing client base relies on Technavios comprehensive coverage, extensive research, and actionable market insights to identify opportunities in existing and potential markets and assess their competitive positions within changing market scenarios.

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COVID-19 Impact and Recovery Analysis | Global Cell Isolation Market 2020-2024 | Evolving Opportunities with Agilent Technologies Inc. and Becton,...

Stem Cell Alopecia Treatment Market Detailed Analysis of Current Industry Figures With Forecasts Growth by 2026 – 3rd Watch News

Stem Cell Alopecia Treatment Market Trends, Size, Shares, Growth, Top Companies, Development, Application, Importance, Overview with Detailed Analysis, Production, Supply, Revenue, Regional Outlook, Status, Competitive Landscape, Future Forecast, Type and End-User, Opportunity, Demand, Historical Data, Business Insights, Research Methodology and many more

Due to the pandemic, we have included a special section on the Impact of COVID 19 on the Stem Cell Alopecia Treatment Market which would mention How the Covid-19 is Affecting the Stem Cell Alopecia Treatment Industry, Market Trends and Potential Opportunities in the COVID-19 Landscape, Covid-19 Impact on Key Regions and Proposal for Stem Cell Alopecia Treatment Players to Combat Covid-19 Impact.

Stem Cell Alopecia Treatment research study involved the extensive usage of both primary and secondary data sources. The research process involved the study of various factors affecting the industry, including the government policy, market environment, competitive landscape, historical data, present trends in the market, technological innovation, upcoming technologies and the technical progress in related industry, and market risks, opportunities, market barriers, and challenges.

Top Key Players of the Global Stem Cell Alopecia Treatment Market:APEX Biologix, Belgravia Center, Kerastem, Riken Research Institute, RepliCel, Sanford Burnham Prebys Medical Discovery Institute,

Download Full PDF Sample Copy of Stem Cell Alopecia Treatment Report: (Including Full TOC, List of Tables & Figures, Chart) : https://www.qyresearch.com/sample-form/form/1491764/global-stem-cell-alopecia-treatment-market

Major manufacturers & their revenues, percentage splits, market shares, growth rates and breakdowns of the product markets are determined through secondary sources and verified through the primary sources.

* Top Key Company Profiles.* Main Business and Rival Information* SWOT Analysis and PESTEL Analysis* Production, Sales, Revenue, Price and Gross Margin* Market Share and Size

Global Stem Cell Alopecia Treatment Market is estimated to reach xxx million USD in 2019 and projected to grow at the CAGR of xx% during 2019-2025. According to the latest report added to the online repository of QY Research the Stem Cell Alopecia Treatment market has witnessed an unprecedented growth till 2019. The extrapolated future growth is expected to continue at higher rates by 2025.

Stem Cell Alopecia Treatment Market Size Estimation

Top-down and bottom-up approaches are used to estimate and validate the global market size for company, regional division, product type and application (end users).

The market estimations in this report are based on the selling price (excluding any discounts provided by the manufacturer, distributor, wholesaler or traders). Market share analysis, assigned to each of the segments and regions are achieved through product utilization rate and average selling price.

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Critical questions addressed by the Stem Cell Alopecia Treatment Market report

What are the key market drivers and restraints?What will be the market size until the end of the forecast period?Which segment is expected to take the lions share?Which region will lead the global Stem Cell Alopecia Treatment market in terms of growth?What will be the key strategies adopted by market leaders in future?What are the upcoming applications?How will the global Stem Cell Alopecia Treatment market develop in the mid to long term?

Table of Contents

1 Stem Cell Alopecia Treatment Market Overview1.1 Stem Cell Alopecia Treatment Product Overview1.2 Stem Cell Alopecia Treatment Market Segment by Type1.2.1 Cloud Service Orchestration1.2.2 API Management1.2.3 Application Integration1.2.4 B2B and Cloud Integration1.2.5 Data Integration1.3 Global Stem Cell Alopecia Treatment Market Size by Type1.3.1 Global Stem Cell Alopecia Treatment Sales and Growth by Type1.3.2 Global Stem Cell Alopecia Treatment Sales and Market Share by Type (2014-2019)1.3.3 Global Stem Cell Alopecia Treatment Revenue and Market Share by Type (2014-2019)1.3.4 Global Stem Cell Alopecia Treatment Price by Type (2014-2019)

2 Global Stem Cell Alopecia Treatment Market Competition by Company2.1 Global Stem Cell Alopecia Treatment Sales and Market Share by Company (2014-2019)2.2 Global Stem Cell Alopecia Treatment Revenue and Share by Company (2014-2019)2.3 Global Stem Cell Alopecia Treatment Price by Company (2014-2019)2.4 Global Top Players Stem Cell Alopecia Treatment Manufacturing Base Distribution, Sales Area, Product Types2.5 Stem Cell Alopecia Treatment Market Competitive Situation and Trends2.5.1 Stem Cell Alopecia Treatment Market Concentration Rate2.5.2 Global Stem Cell Alopecia Treatment Market Share of Top 5 and Top 10 Players2.5.3 Mergers & Acquisitions, Expansion

3 Stem Cell Alopecia Treatment Company Profiles and Sales Data3.1 IBM3.1.1 Company Basic Information, Manufacturing Base and Competitors3.1.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.1.3 IBM Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.1.4 Main Business Overview3.2 Microsoft3.2.1 Company Basic Information, Manufacturing Base and Competitors3.2.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.2.3 Microsoft Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.2.4 Main Business Overview3.3 Oracle3.3.1 Company Basic Information, Manufacturing Base and Competitors3.3.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.3.3 Oracle Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.3.4 Main Business Overview3.4 Software3.4.1 Company Basic Information, Manufacturing Base and Competitors3.4.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.4.3 Software Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.4.4 Main Business Overview3.5 TIBCO Software3.5.1 Company Basic Information, Manufacturing Base and Competitors3.5.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.5.3 TIBCO Software Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.5.4 Main Business Overview3.6 Accenture3.6.1 Company Basic Information, Manufacturing Base and Competitors3.6.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.6.3 Accenture Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.6.4 Main Business Overview3.7 Adeptia3.7.1 Company Basic Information, Manufacturing Base and Competitors3.7.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.7.3 Adeptia Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.7.4 Main Business Overview3.8 Atos3.8.1 Company Basic Information, Manufacturing Base and Competitors3.8.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.8.3 Atos Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.8.4 Main Business Overview3.9 Axway3.9.1 Company Basic Information, Manufacturing Base and Competitors3.9.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.9.3 Axway Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.9.4 Main Business Overview3.10 BT Global Services3.10.1 Company Basic Information, Manufacturing Base and Competitors3.10.2 Stem Cell Alopecia Treatment Product Category, Application and Specification3.10.3 BT Global Services Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin(2014-2019)3.10.4 Main Business Overview

4 Stem Cell Alopecia Treatment Market Status and Outlook by Regions4.1 Global Market Status and Outlook by Regions4.1.1 Global Stem Cell Alopecia Treatment Market Size and CAGR by Regions4.1.2 North America4.1.3 Asia-Pacific4.1.4 Europe4.1.5 South America4.1.6 Middle East and Africa4.2 Global Stem Cell Alopecia Treatment Sales and Revenue by Regions4.2.1 Global Stem Cell Alopecia Treatment Sales and Market Share by Regions (2014-2019)4.2.2 Global Stem Cell Alopecia Treatment Revenue and Market Share by Regions (2014-2019)4.2.3 Global Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin (2014-2019)4.3 North America Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin4.3.1 United States4.3.2 Canada4.3.3 Mexico4.4 Europe Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin4.4.1 Germany4.4.2 UK4.4.3 France4.4.4 Italy4.4.5 Russia4.4.6 Turkey4.5 Asia-Pacific Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin4.5.1 China4.5.2 Japan4.5.3 Korea4.5.4 Southeast Asia4.5.4.1 Indonesia4.5.4.2 Thailand4.5.4.3 Malaysia4.5.4.4 Philippines4.5.4.5 Vietnam4.5.5 India4.5.6 Australia4.6 South America Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin4.6.1 Brazil4.7 Middle East and Africa Stem Cell Alopecia Treatment Sales, Revenue, Price and Gross Margin4.7.1 Egypt4.7.2 GCC Countries

5 Stem Cell Alopecia Treatment Application/End Users5.1 Stem Cell Alopecia Treatment Segment by Application5.1.1 Banking, Financial Services, and Insurance5.1.2 Consumer Goods and Retail5.1.3 Education5.1.4 Government and Public Sector5.1.5 Healthcare and Life Sciences5.1.6 Manufacturing5.1.7 Media and Entertainment5.1.8 Telecommunication and Ites5.1.9 Others5.2 Global Stem Cell Alopecia Treatment Product Segment by Application5.2.1 Global Stem Cell Alopecia Treatment Sales by Application5.2.2 Global Stem Cell Alopecia Treatment Sales and Market Share by Application (2014-2019)

6 Global Stem Cell Alopecia Treatment Market Forecast6.1 Global Stem Cell Alopecia Treatment Sales, Revenue Forecast (2019-2025)6.1.1 Global Stem Cell Alopecia Treatment Sales and Growth Rate Forecast (2019-2025)6.1.1 Global Stem Cell Alopecia Treatment Revenue and Growth Rate Forecast (2019-2025)6.2 Global Stem Cell Alopecia Treatment Forecast by Regions6.2.1 North America Stem Cell Alopecia Treatment Sales and Revenue Forecast (2019-2025)6.2.2 Europe Stem Cell Alopecia Treatment Sales and Revenue Forecast (2019-2025)6.2.3 Asia-Pacific Stem Cell Alopecia Treatment Sales and Revenue Forecast (2019-2025)6.2.3.1 China6.2.3.2 Japan6.2.3.3 Korea6.2.3.4 Southeast Asia6.2.3.5 India6.2.3.6 Australia6.2.4 South America Stem Cell Alopecia Treatment Sales and Revenue Forecast (2019-2025)6.2.5 Middle East and Africa Stem Cell Alopecia Treatment Sales and Revenue Forecast (2019-2025)6.2.5.1 Egypt6.2.5.2 GCC Countries6.3 Stem Cell Alopecia Treatment Forecast by Type6.3.1 Global Stem Cell Alopecia Treatment Sales and Revenue Forecast by Type (2019-2025)6.3.2 Cloud Service Orchestration Gowth Forecast6.3.3 API Management Gowth Forecast6.4 Stem Cell Alopecia Treatment Forecast by Application6.4.1 Global Stem Cell Alopecia Treatment Sales Forecast by Application (2019-2025)6.4.2 Global Stem Cell Alopecia Treatment Forecast in Banking, Financial Services, and Insurance6.4.3 Global Stem Cell Alopecia Treatment Forecast in Consumer Goods and Retail

7 Stem Cell Alopecia Treatment Upstream Raw Materials7.1 Stem Cell Alopecia Treatment Key Raw Materials7.1.1 Key Raw Materials7.1.2 Key Raw Materials Price7.1.3 Raw Materials Key Suppliers7.2 Manufacturing Cost Structure7.2.1 Raw Materials7.2.2 Labor Cost7.2.3 Manufacturing Expenses7.3 Stem Cell Alopecia Treatment Industrial Chain Analysis

8 Marketing Strategy Analysis, Distributors8.1 Marketing Channel8.1.1 Direct Marketing8.1.2 Indirect Marketing8.1.3 Marketing Channel Development Trend8.2 Distributors8.3 Downstream Customers

9 Research Findings and Conclusion

AppendixMethodology/Research ApproachResearch Programs/DesignMarket Size EstimationMarket Breakdown and Data TriangulationData SourceSecondary SourcesPrimary SourcesDisclaimer

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Stem Cell Alopecia Treatment Market Detailed Analysis of Current Industry Figures With Forecasts Growth by 2026 - 3rd Watch News