China to conduct first human trials on embryonic stem cells – BSA bureau (press release)

This will mark the start of the first clinical trial in China using human embryonic stem (ES) cells, and the first one worldwide aimed at treating Parkinsons disease using ES cells from fertilized embryos.

Surgeons in the Chinese city of Zhengzhou will begin the process of injecting the brains of people with Parkinsons disease with 4 million immature neurons derived from human embryonic stem cells, writes a leading science news website.

This will mark the start of the first clinical trial in China using human embryonic stem (ES) cells, and the first one worldwide aimed at treating Parkinsons disease using ES cells from fertilized embryos. In a second trial starting around the same time, a different team in Zhengzhou will use ES cells to target vision loss caused by age-related macular degeneration.

The experiments will represent the first clinical trials of ES cells under regulations that China adopted in 2015, in an attempt to ensure the ethical and safe use of stem cells in the clinic. China previously had no clear regulatory framework, and many companies had used that gap as an excuse to market unproven stem cell treatments.

In the first, the surgeons will inject ES-cell-derived neuronal precursor cells into the brains of individuals with Parkinsons disease. The only previous trial using ES cells to treat Parkinsons began last year in Australia; participants there received stem cells from parthenogenetic embryos, unfertilized eggs that are triggered in the lab to start embryonic development.

Stem cell specialist at the Chinese Academy of Sciences Institute of Zoology in Beijing, Qi Zhou is responsible for taking the lead in both trials.

However, the announcement of the trials brings with it concerns. Jeanne Loring, a stem cell biologist at the Scripps Research Institute in La Jolla, California, who is also planning stem cell trials for Parkinsons, is concerned that the Australian and Chinese trials use neural precursors and not ES-cell-derived cells that have fully committed to becoming dopamine-producing cells. Precursor cells can turn into other kinds of neurons, and could accumulate dangerous mutations during their many divisions, explains Loring to a news website.

Despite this, the Zhou and the Australian team defend their choices as the animal trials conducted thus far have been promising. If Zhou and the rest of the team is correct, this will represent a major step forward for the entire world and usher in a new era ofstem cellresearch.

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China to conduct first human trials on embryonic stem cells - BSA bureau (press release)

Embryonic stem cell trials to launch in China – BioEdge

China will begin trialling the use of embryonic stem-cells (ES) to treat Parkinsons disease and macular degeneration, in a move that has met with criticism from international experts.

The trials, which come in the wake of new stem-cell regulations introduced in China in 2015, will test the efficacy of injecting ES-derived cells into damaged areas of the brain and eyes.

In one trial, ES-derived neuronal-precursor cells will be injected into the areas of the brain affected by Parkinsons disease in attempt to regenerate dopamine-producing tissue. In another trial, the ES-derived retinal cells will be injected into eyes of people with age related macular degeneration. It is believed that the retinal cells may be able to replace cells damaged as a result of epithelial tissue degeneration.

It will be a major new direction for China, Pei Xuetao, a stem-cell scientist at the Beijing Institute of Transfusion Medicine who is on the central-government committee that approved the trials, told Nature.

Other researchers who work on Parkinsons disease, however, worry that the trials might be misguided.

Jeanne Loring, a stem-cell biologist at the Scripps Research Institute in La Jolla, California, who is also planning stem-cell trials for Parkinsons, is concerned that the Chinese trials use neural precursors and not ES-cell-derived cells that have fully committed to becoming dopamine-producing cells. Precursor cells can turn into other kinds of neurons, and could accumulate dangerous mutations during their many divisions, says Loring. Not knowing what the cells will become is troubling.

Lorenz Studer, a stem-cell biologist at the Memorial Sloan Kettering Cancer Center in New York City, says that support is not very strong for the use of precursor cells. I am somewhat surprised and concerned, as I have not seen any peer-reviewed preclinical data on this approach, he told Nature.

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Embryonic stem cell trials to launch in China - BioEdge

Trials of Embryonic Stem Cells to Launch in China

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In the next few months, surgeons in the Chinese city of Zhengzhou will carefully drill through the skulls of people with Parkinsons disease and inject 4 million immature neurons derived from human embryonic stem cells into their brains. Then they will patch the patients up, send them home and wait.

This will mark the start of the first clinical trial in China using human embryonic stem (ES) cells, and the first one worldwide aimed at treating Parkinsons disease using ES cells from fertilized embryos. In a second trial starting around the same time, a different team in Zhengzhou will use ES cells to target vision loss caused by age-related macular degeneration.

The experiments will also represent the first clinical trials of ES cells under regulations that China adopted in 2015, in an attempt to ensure the ethical and safe use of stem cells in the clinic. China previously had no clear regulatory framework, and many companies had used that gap as an excuse to market unproven stem-cell treatments.

It will be a major new direction for China, says Pei Xuetao, a stem-cell scientist at the Beijing Institute of Transfusion Medicine who is on the central-government committee that approved the trials. Other researchers who work on Parkinsons disease, however, worry that the trials might be misguided.

Both studies will take place at the First Affiliated Hospital ofZhengzhouUniversity in Henan province. In the first, surgeons will inject ES-cell-derived neuronal-precursor cells into the brains of individuals with Parkinsons disease. The only previous trial using ES cells to treat Parkinsons began last year in Australia; participants there received stem cells from parthenogenetic embryosunfertilized eggs that are triggered in the lab to start embryonic development.

In the other Zhengzhou trial, surgeons will take retinal cells derived from ES cells and transplant them into the eyes of people with age-related macular degeneration. The team will follow a similar procedure to that of previous ES-cell trials carried out by researchers in the United States and South Korea.

Qi Zhou, a stem-cell specialist at the Chinese Academy of Sciences Institute of Zoology in Beijing, is leading both efforts. For the Parkinsons trial, his team assessed hundreds of candidates and have so far have picked ten who best match the ES cells in the cell bank, to reduce the risk of the patients bodies rejecting the cells.

The 2015 regulations state that hospitals planning to carry out stem-cell clinical work must use government-certified ES-cell lines and pass hospital-review procedures. Zhous team completed four years of work with a monkey model of Parkinsons, and has met the government requirements, he says.

Parkinsons disease is caused by a deficit in dopamine produced by brain cells. Zhous team will coax ES cells to develop into precursors to neurons, and will then inject them into the striatum, a central region of the brain implicated in the disease.

In their unpublished study of 15 monkeys, the researchers did not observe any improvements in movement at first, says Zhou. But at the end of the first year, the team examined the brains of half the monkeys and found that the stem cells had turned into dopamine-releasing cells. He says that they saw 50% improvement in the remaining monkeys over the next several years. We have all the imaging data, behavioural data and molecular data to support efficacy, he says. They are preparing a publication, but Zhou says that they wanted to collect a full five years worth of animal data.

Jeanne Loring, a stem-cell biologist at the Scripps Research Institute in La Jolla, California, who is also planning stem-cell trials for Parkinsons, is concerned that the Australian and Chinese trials use neural precursors and not ES-cell-derived cells that have fully committed to becoming dopamine-producing cells. Precursor cells can turn into other kinds of neurons, and could accumulate dangerous mutations during their many divisions, says Loring. Not knowing what the cells will become is troubling.

But Zhou and the Australian team defend their choices. Russell Kern, chief scientific officer of the International Stem Cell Corporation in Carlsbad, California, which is providing the cells for and managing the Australian trial, says that in preclinical work, 97% of them became dopamine-releasing cells.

Lorenz Studer, a stem-cell biologist at the Memorial Sloan Kettering Cancer Center in New York City who has spent years characterizing such neurons ahead of his own planned clinical trials, says that support is not very strong for the use of precursor cells. I am somewhat surprised and concerned, as I have not seen any peer-reviewed preclinical data on this approach, he says.

Studers and Lorings teams are part of an international consortium that coordinates stem-cell treatments for Parkinsons. In the next two years, five groups in the consortium plan to run trials using cells fully committed to becoming dopamine-producing cells.

Regenerative neurobiologist Malin Parmar, who heads one of the teams at Lund University in Sweden, says that the groups are all rapidly moving towards clinical trials, and this field will be very exciting in the coming years.

Source & Credits: ScientificAmerican

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Trials of Embryonic Stem Cells to Launch in China

Trials of embryonic stem cells to launch in China : Nature …

Jason Lee/Reuters

Former Chinese leader Deng Xiaoping had Parkinsons disease, one of the first targets of embryonic-stem-cell therapies being tested in China.

In the next few months, surgeons in the Chinese city of Zhengzhou will carefully drill through the skulls of people with Parkinsons disease and inject 4 million immature neurons derived from human embryonic stem cells into their brains. Then they will patch the patients up, send them home and wait.

This will mark the start of the first clinical trial in China using human embryonic stem (ES) cells, and the first one worldwide aimed at treating Parkinsons disease using ES cells from fertilized embryos. In a second trial starting around the same time, a different team in Zhengzhou will use ES cells to target vision loss caused by age-related macular degeneration.

The experiments will also represent the first clinical trials of ES cells under regulations that China adopted in 2015, in an attempt to ensure the ethical and safe use of stem cells in the clinic. China previously had no clear regulatory framework, and many companies had used that gap as an excuse to market unproven stem-cell treatments.

It will be a major new direction for China, says Pei Xuetao, a stem-cell scientist at the Beijing Institute of Transfusion Medicine who is on the central-government committee that approved the trials. Other researchers who work on Parkinsons disease, however, worry that the trials might be misguided.

Both studies will take place at the First Affiliated Hospital ofZhengzhouUniversity in Henan province. In the first, surgeons will inject ES-cell-derived neuronal-precursor cells into the brains of individuals with Parkinsons disease. The only previous trial using ES cells to treat Parkinsons began last year in Australia; participants there received stem cells from parthenogenetic embryosunfertilized eggs that are triggered in the lab to start embryonic development.

In the other Zhengzhou trial, surgeons will take retinal cells derived from ES cells and transplant them into the eyes of people with age-related macular degeneration. The team will follow a similar procedure to that of previous ES-cell trials carried out by researchers in the United States and South Korea.

Qi Zhou, a stem-cell specialist at the Chinese Academy of Sciences Institute of Zoology in Beijing, is leading both efforts. For the Parkinsons trial, his team assessed hundreds of candidates and have so far have picked ten who best match the ES cells in the cell bank, to reduce the risk of the patients bodies rejecting the cells.

The 2015 regulations state that hospitals planning to carry out stem-cell clinical work must use government-certified ES-cell lines and pass hospital-review procedures. Zhous team completed four years of work with a monkey model of Parkinsons, and has met the government requirements, he says.

Parkinsons disease is caused by a deficit in dopamine produced by brain cells. Zhous team will coax ES cells to develop into precursors to neurons, and will then inject them into the striatum, a central region of the brain implicated in the disease.

In their unpublished study of 15 monkeys, the researchers did not observe any improvements in movement at first, says Zhou. But at the end of the first year, the team examined the brains of half the monkeys and found that the stem cells had turned into dopamine-releasing cells. He says that they saw 50% improvement in the remaining monkeys over the next several years. We have all the imaging data, behavioural data and molecular data to support efficacy, he says. They are preparing a publication, but Zhou says that they wanted to collect a full five years worth of animal data.

Jeanne Loring, a stem-cell biologist at the Scripps Research Institute in La Jolla, California, who is also planning stem-cell trials for Parkinsons, is concerned that the Australian and Chinese trials use neural precursors and not ES-cell-derived cells that have fully committed to becoming dopamine-producing cells. Precursor cells can turn into other kinds of neurons, and could accumulate dangerous mutations during their many divisions, says Loring. Not knowing what the cells will become is troubling.

But Zhou and the Australian team defend their choices. Russell Kern, chief scientific officer of the International Stem Cell Corporation in Carlsbad, California, which is providing the cells for and managing the Australian trial, says that in preclinical work, 97% of them became dopamine-releasing cells.

Lorenz Studer, a stem-cell biologist at the Memorial Sloan Kettering Cancer Center in New York City who has spent years characterizing such neurons ahead of his own planned clinical trials, says that support is not very strong for the use of precursor cells. I am somewhat surprised and concerned, as I have not seen any peer-reviewed preclinical data on this approach, he says.

Studers and Lorings teams are part of an international consortium that coordinates stem-cell treatments for Parkinsons. In the next two years, five groups in the consortium plan to run trials using cells fully committed to becoming dopamine-producing cells.

Regenerative neurobiologist Malin Parmar, who heads one of the teams at Lund University in Sweden, says that the groups are all rapidly moving towards clinical trials, and this field will be very exciting in the coming years.

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Trials of embryonic stem cells to launch in China : Nature ...

From baby mice to designer humans? – Brnow

Scientists in Japan are growing mice from skin cells. A team of researchers at Kyushu University last year converted tail cells from adult female mice into viable eggs, and then inseminated those eggs to produce embryos. They implanted the embryos in female mice who gave birth to healthy baby mice. The process, called in vitro gametogenesis (IVG), is a big leap from todays in vitro fertilization (IVF). With IVG, doctors can artificially create eggs and sperm by coaxing cells from other parts of the body into stem cells, and then into eggs and sperm. Researchers say it is only a matter of time before they can use the process for human reproduction. But experts warn of serious ethical, medical and legal consequences for using this new technology on humans. In a cautionary article published earlier this year in the journal Science Translational Medicine, a group of academics from Harvard and Brown universities noted the technology promises to transform the fields of reproductive and regenerative medicine, but also creates vast ethical and social policy challenges that must be addressed. With IVG, creating life no longer would require a man and a woman: Two men could make a baby biologically related to them both using the skin cells of one to make an egg, and the sperm of the other. A woman could make a baby by herself using her cell-turned-sperm and her egg, almost like cloning. A group of three or four people could create a baby by creating two embryos, and then taking an egg from one and a sperm from the other, creating another embryo with multiple parents. Such scenarios inevitably would affect the traditional understanding of parenting. The article also addressed the potential for unauthorized use of biomaterials: Someone retrieves a skin cell from a hotel room bed or bathroom, creating a baby biologically related to someone without their knowledge. Should the law criminalize such an action? If it takes place, should the law consider the source of the skin cells to be a legal parent to the child, or should it distinguish an individuals genetic and legal parentage? the authors wrote. They also raised the potential for bioethical issues on a massive scale. IVG may raise the specter of embryo farming on a scale currently unimagined, which might exacerbate concerns about the devaluation of human life, wrote the authors, pointing to the inevitable destruction of large numbers of embryos, the commercialization of egg production, the creation of an all but inexhaustible supply of embryonic stem cells for research and the open invitation for a couple to create designer babies due to limitless eggs. But significant scientific hurdles remain. People are a lot more complicated than mice, Susan Solomon, chief executive of the New York Stem Cell Foundation, told The New York Times. And weve often seen that the closer you get to something, the more obstacles you discover. Despite those hurdles, the articles authors warn IVG technology is moving faster than our conversations about the ethical questions it raises. We have come to realize that scientific developments are outpacing our ability to think through them, Eli Y. Adashi, a medical science professor at Brown, told The New York Times. Its a challenge for which we are not fully prepared. (EDITORS NOTE Kiley Crossland writes for WORLD News Service, a division of WORLD Magazine, worldmag.com, based in Asheville, N.C. Used by permission.)

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From baby mice to designer humans? - Brnow

World-first trials have been launched to treat Parkinson’s and blindness with embryonic stem cells – ScienceAlert

In a world first, surgeons in the Chinese city of Zhengzhou are planning to inject stem cells derived from human embryos into the brains of patients with Parkinson's disease with the aim of treating their debilitating symptoms.

Meanwhile, another medical team in the same city is aiming to target vision loss using embryonic stem cells (ESC) to replace lost cells in the retina, marking a new direction in China in the wake of major changes in how the country regulates stem cell treatments.

While similar treatments on Parkinson's patients have already been tested in Australia, those trials relied on cells taken from eggs that were forced to divide without first being fertilised in an effort to circumvent any ethical concerns.

Stem cells are a little like blank slates that are yet to take on a specific task. If you rewind the clock on any of your body's tissues, its cells will become less specialised, until you're left with a cell with a lot of potential to become nearly anything.

In the case of both kinds of embryonic stem cells, divided egg cells are subjected to various treatments to encourage them to develop into replacement cells that could treat a condition in a recipient.

The symptoms of Parkinson's disease are largely caused by a loss of nervous tissue deep inside the brain in an area called the basal ganglia.

Losing those cells means a loss of a neurotransmitter called dopamine, and with it a lower ability to control nervous impulses that would prevent muscles in the extremities from activating.

In the case of a condition called macular degeneration, damage to a layer of tissue called the retinal pigment epithelium at the back of the eye causes the light-catching cells above it to die.

By turning ESC into cells that can naturally develop into the tissues that have deteriorated such as the precursors to neurons that can produce dopamine, or into retinal tissue and then injecting it into the target site, the researchers hope to improve the lost functions.

Not everybody is convinced of the success of trials such as those being done in China and last year in Australia.

A stem cell biologist from the Scripps Research Institute in California, Jeanne Loring, believes the choice of cell used in both Parkinson's disease trials won't be specialised enough to match expected results.

"Not knowing what the cells will become is troubling," Loring told David Cyranoski at Nature.

But the research team in China remains confident in its decision.

Qi Zhou from the Chinese Academy of Sciences Institute of Zoology in Beijing is the stem cell specialist leading both sets of ESC trials, and says four years of animal trials conducted on monkeys have so far showed promising results.

"We have all the imaging data, behavioural data, and molecular data to support efficacy," Zhou told Nature.

He also claims the team conducting the Parkinson's trial have been selective with their potential candidates, choosing patients who will have the least chance of rejecting the ESCs from the cell bank.

In 2015, China introduced tough new regulations to deal with the growing problem of 'rogue clinics' offering stem cell treatments without due record keeping or process, making it hard to evaluate safety, or even the types of cells used in the treatments.

The changes are set to improve the ethics and safety of stem cell treatments by enforcing the use of cells through a regulatory body, ensuring informed patient consent, and permitting treatments only through authorised hospitals.

Time will tell if the regulations can be enforced, but for stem cell researchers, the changes are positive.

"It will be a major new direction for China," stem cell scientist Pei Xuetaotold Nature.

If the results are as good as the teams in Australia and China predict, it could also set new standards for the world.

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World-first trials have been launched to treat Parkinson's and blindness with embryonic stem cells - ScienceAlert

Scientists find that smoking harms livers of unborn babies – BBC News

The National

Scientists find that smoking harms livers of unborn babies BBC News Scientists found that the cocktail of chemicals in cigarettes is particularly harmful to developing liver cells. They developed a method of studying the effects of maternal smoking on liver tissue using embryonic stem cells. The team, led by the ... Smoking by pregnant women damages the liver cells of their unborn babiesThe National all 4 news articles »

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Scientists find that smoking harms livers of unborn babies - BBC News

A new method for creating safer induced pluripotent stem cells – Phys.Org

May 24, 2017 Journal cover image. Credit: Future Science Group

Induced pluripotent stem cells (IPSCs) hold great promise in regenerative medicine, personalized medicine and drug discovery. However, while avoiding the ethical controversies associated with embryonic stem cells, they carry neoplastic risk owing to the use of the oncogenes c-Myc and Lin28. This has limited their utility in the biomedical arena.

Work has previously demonstrated that IPSC generation can be uncoupled from c-Myc, but until now a viable oncogene- and virus-free method has proven elusive.

A new research article from Alan B Moy and colleagues at Cellular Engineering Technologies (IA, USA) and The John Paul II Medical Research Institute (IA, USA) describes a promising new IPSC reprogramming approach that attempts to solve these issues. The open access article, "Efficient method to create integration-free, virus-free, Myc and Lin28-free human induced pluripotent stem cells from adherent cells", is available in Future Science OA.

Their approach saw adherent fibroblasts reprogrammed using a combination of reprogramming molecules and episomal vectors. The combinatorial approach was successful, yielding colonies in which 100% expressed SSEA4.

"Our reprogramming method provides patient- and disease-specific IPSC[s] for drug discovery and personalized medicine applications with lower risk of oncogenic perturbations due to Lin28 and Myc", noted the authors. "The reprogramming method paves a pathway for autologous and allogeneic cell therapy that satisfies regulatory requirements."

The team anticipates that virus- and oncogene-free IPSCs could advance cell therapies, diagnostics and personalized medicine. Furthermore, they envision the technology helping to reduce clinical trial failure rates and improve drug development.

Explore further: Stem cell reprogramming factor controls change in cellular energy generation

More information: Anant Kamath et al, Efficient method to create integration-free, virus-free,and-free human induced pluripotent stem cells from adherent cells, Future Science OA (2017). DOI: 10.4155/fsoa-2017-0028

Provided by: Future Science Group

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The potential to use a patient's own cells to treat non-healing chronic wounds - a serious complication of diabetes - took an important step forward as researchers successfully reprogrammed skin cells taken from diabetic ...

It's been more than 10 years since Japanese researchers Shinya Yamanaka, M.D., Ph.D., and his graduate student Kazutoshi Takahashi, Ph.D., developed the breakthrough technique to return any adult cell to its earliest stage ...

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How do you improve a Nobel Prize-winning discovery? Add a debilitating disease-causing gene mutation.

Scientists have discovered the gene essential for chemically reprogramming human amniotic stem cells into a more versatile state similar to embryonic stem cells, in research led by UCL and Heinrich Heine University.

There are significant gaps in our knowledge on the evolution of sex, according to a research review on sex chromosomes from Lund University in Sweden. Even after more than a century of study, researchers do not know enough ...

(Phys.org)Eusocial insects are predominantly dependent on chemosensory communication to coordinate social organization and define group membership. As the social complexity of a species increases, individual members require ...

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A new method for creating safer induced pluripotent stem cells - Phys.Org

Republicans Want Trump to Fire NIH Director Who Supports Embryonic Stem Cell Research – LifeNews.com

Republicans in Congress are urging President Donald Trump to oust the director of the National Institutes of Health because of his support for embryonic stem cell research that involves the destruction of human life.

40 Republicans in the House of Representatives wrote president Trump urging him to get rid of NIH director Francis Collins because of his support for the practice, which is opposed by pro-life organizations.

While pro-life advocates strongly support scientific research, they oppose embryonic stem cell research because the only way to obtain embryonic stem cells is to destroy unique human beings just days after conception. On the other hand, ethical adult stem cell research has produced cures or treatments for well over 100 diseases or medical conditions and involves no destruction of human life. Embryonic stem cell research has still yet to treat a single human being successfully.

Here is more from Politico:

Forty House Republicans are urging President Donald Trump to fire the director of the NIH over his support for embryonic and stem cell research that they say conflicts with Trumps pro-life direction.

The Republican House members, in a letter led by Rep. Jim Banks (R-Ind.), question NIH Director Francis Collins support for embryonic cloning and for stem cell research that involves the destruction of human embryos.

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While we deeply respect Dr. Collins Christian faith and commitment to public service, the stances that Dr. Collins has taken in the past regarding embryonic stem cell research and human cloning are not life-affirming and directly conflict with the pro-life direction of your new presidency, the GOP lawmakers wrote. It is because of this troubling paradox that we ask you to re-consider his leadership role at NIH.

Collins has led the NIH since 2009, when he was unanimously confirmed by the Senate.

It has been unclear whether Trump will keep Collins, who came in at the beginning of the Obama administration, at the helm of the biomedical research institution. But Collins has wide support from both Republicans and Democrats.

In fact, several top Republicans former House Energy and Commerce Chairman Fred Upton, Senate HELP Chairman Lamar Alexander and key health care appropriators such as Sen. Roy Blunt and Rep. Tom Cole asked Trump in December to keep Collins.

But the 40 Republicans argue Collins doesnt share in their partys position on embryonic research. They wrote that Collins stance is particularly disturbing considering that NIHs funding for human embryonic stem cell research increased from $146 million in 2012 to $180 million in 2015.

The Obama administration forced Americans to pay for embryonic stem cell research involving the destruction of human life. National Institutes of Health chief Francis Collins approved taxpayer funding of dozens of lines of embryonic stem cells. The cells can only be obtained by destroying unborn children days after conception at which point human embryos are unique human beings.

Obama issued an executive order overturning the limits President Bush put in place on any new embryonic stem cell research funding. Bush directed federal dollars mostly to adult stem cells that are already helping patients now.

The Family Research Council responded to the Obama administrations move by saying that adult stem cells are already helping diabetes patients.

Of course, when it comes to juvenile (type I) diabetes, adult stem cells have already shown success at treating diabetes patients, the pro-life group noted.

It added, Interestingly, none of the human embryonic stem cell lines approved thus far are from the original group of 21 lines that had been receiving NIH funding, only one (H1one of the original five Thomson lines) has been submitted for approval, with only two other lines from Cellartis supposedly to be submitted for review.

This is surprising given that the vast majority of human embryonic stem cell research has been done with those previous lines, thus forcing most embryonic stem cell researchers to start over on experiments with new lines the group continued.

It also shows that this gold rush is just that, an attempt to grab more money built on embryo destruction, not built on science, FRC concluded.

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Republicans Want Trump to Fire NIH Director Who Supports Embryonic Stem Cell Research - LifeNews.com

ESC-Track: A computer workflow for 4-D segmentation, tracking, lineage tracing and dynamic context analysis of ESCs – BioTechniques.com

ESC-Track: A computer workflow for 4-D segmentation, tracking, lineage tracing and dynamic context analysis of ESCs

Laura Fernndez-de-Manel1, Covadonga Daz-Daz2, Daniel Jimnez-Carretero1, Miguel Torres2, and Mara C. Montoya1

1Cellomics Unit 2Cardiovascular Developmental Program, Cell & Developmental Biology Area, Centro Nacional de Investigaciones Cardiovasculares CNIC, Madrid, Spain

BioTechniques, Vol. 62, No. 5, May 2017, pp. 215222

Abstract

Embryonic stem cells (ESCs) can be established as permanent cell lines, and their potential to differentiate into adult tissues has led to widespread use for studying the mechanisms and dynamics of stem cell differentiation and exploring strategies for tissue repair. Imaging live ESCs during development is now feasible due to advances in optical imaging and engineering of genetically encoded fluorescent reporters; however, a major limitation is the low spatio-temporal resolution of long-term 3-D imaging required for generational and neighboring reconstructions. Here, we present the ESC-Track (ESC-T) workflow, which includes an automated cell and nuclear segmentation and tracking tool for 4-D (3-D + time) confocal image data sets as well as a manual editing tool for visual inspection and error correction. ESC-T automatically identifies cell divisions and membrane contacts for lineage tree and neighborhood reconstruction and computes quantitative features from individual cell entities, enabling analysis of fluorescence signal dynamics and tracking of cell morphology and motion. We use ESC-T to examine Myc intensity fluctuations in the context of mouse ESC (mESC) lineage and neighborhood relationships. ESC-T is a powerful tool for evaluation of the genealogical and microenvironmental cues that maintain ESC fitness.

Stem cells provide essential functions during embryonic development and tissue regeneration. Mouse embryonic stem cells (mESCs) are derived from pluripotent cells of the early mouse embryo and can be maintained as stable cell lines with a high self-renewal capacity. They provide a versatile in vitro model for understanding differentiation of human tissues, and their study has led to major advances in cell and developmental biology (1,2). A key challenge in the field is to understand the mechanisms involved in guiding stem cell fate (3-5), which have broad applications in biomedicine, from elucidating the causes of cancer to the use of stem cells in regenerative medicine. Thus, the biological properties of ESCs are a matter of great scientific, commercial, and medical interest.

ESC-Track (ESC-T) is a computational tool for automated segmentation and tracking of single mouse embryonic stem cells (mESCs) from live-cell 4-D confocal image data sets. The ESC-T workflow enables the extraction of parameters related to fluorescence signal localization and dynamics, cellular morphology, and cell motion for individual cells in the context of lineage and neighborhood relationships.

Optical imaging advances have led to the emergence of powerful live imaging tools with individual cell resolution in three-dimensional (3-D) space and in time (3-D + time or 4-D) (6,7). Moreover, a new generation of fluorescent proteins and dyes allows biochemical characterization of signaling pathways in intact living cells (8). Tagging by fluorescent proteins enables positional tracking of any given cell over time, which is easily achieved when the population of tagged cells is distributed among non-expressing cells by virtue of lineage or in experimental mosaics, but it becomes challenging when a fluorescent protein label is widely expressed (9). The ability to track and analyze live cells in time-lapse 4-D microscopy images is a matter of intense research (10,11) since visual inspection and analysis are insufficient to extract meaningful insights, making automated tracking and quantitative analysis of cells an absolute requirement. This is such a challenging task that several competitions have been carried out in order to evaluate cell segmentation and tracking algorithms (12,13). Computational tools are essential for extracting quantitative measurements from stem cell populations growing in 3-D physiological conditions and to translate the measurements into biological knowledge, allowing the study of a range of cell behaviors, such as motility, cell division, death, phagocytosis, etc. Most of these methods have been applied to Drosophila (14-18) and zebrafish (19-21) embryogenesis, or plant morphogenesis (22) studies. Of special relevance to the field of stem cell biology is the ability to integrate the cell behavior analysis with information about lineage (parentprogeny) and contextual (neighborhood) cellular relationships (9,11). In the last decade, several generic processing and tracking packages, such as Icy (23), Cell Profiler (24), tTt (25), qTfy (25), or the Fiji plugin TrackMate (26,27) have been reported. Some complex methods have been developed for specific applications, such as MARS (22), ACME (21), EDGE-4-D (17), and RACE (18) for particle (28), nuclear (16,29) or cellular (17,18,20-22) segmentation, and STARRYNITE (29), U-TRACK (28), ALT (22), EDGE-4-D (17), and TGMM (16) for tracking.

Here, we present a computational workflow that allows the automated segmentation and tracking of individual mESCs from live-cell 4-D confocal image data sets based on the combination of membrane and non-homogeneous nuclear signals, allowing lineage and neighborhood reconstruction. The workflow enables the extraction of parameters for fluorescence signal localization and dynamics, cellular morphological characteristics, and motion-related aspects of individual cells in the context of lineage and neighborhood relationships. ESC-T was used to study Myc dynamics in mESC cultures, and it proved to be a very valuable computational tool for stem cell research as it allowed the evaluation of genealogical and microenvironmental cues during mouse ESC culture in an unprecedented manner.

Automatic cell and nuclei segmentation and cell tracking. The proposed pipeline (Figure 1) uses images obtained from ESCs expressing tdTomato and GFP-MYC signals as described in the Supplementary Material. The pre-processing step consists of median filtering combining both nuclear and membrane signals (mycGFP median minus tdTomato median) (Figure 1, Steps 14) and is followed by application of a 2-D watershed segmentation algorithm, rendering 2-D sets (cell portions) (Figure 1, Step 5). Spatiotemporal (3-D + t) association rules based on the overlap of sets are applied to connect sets in 3-D space and time for automatic segmentation and tracking through the following pipeline:

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ESC-Track: A computer workflow for 4-D segmentation, tracking, lineage tracing and dynamic context analysis of ESCs - BioTechniques.com