(A) Nuclear DNA genotyping from four human NT-ESC lines (hESO-NT1, hESO-NT2, hESO-NT3, and hESO-NT4) determined by microsatellite parentage analysis. A total of 24 microsatellite markers were used for each analysis. The representative markers for D2S1333 and D4S413 loci demonstrate that the nuclear DNA in these cell lines was exclusively derived from the somatic HDF-f cell line. No contribution of oocyte nuclear DNA was detected. (B) mtDNA genotyping by Sanger sequencing demonstrated that all NT-ESC lines contain oocyte mtDNA. (C) Cytogenetic G-banding analysis confirmed that all NT-ESCs exhibited a normal 46XX karyotype (hESO-NT1 result is representative). (D) Human NT-ESCs expressed standard pluripotency markers detected by immunocytochemistry for antibodies against OCT4, NANOG, SOX2, SSEA-4, TRA-160, and TRA-181. Original magnification, 200; Ph, phase contrast. Note that the upper-left image for hESO-NT1 is the same shown in Figure 2F. The upper-right image for hESO-7 is the same shown in Figure S5 (upper-right). (E) Histological analysis of teratoma tumors produced after injection of human NT-ESCs into SCID mice. An arrow and arrowhead in the top panel indicate intestinal-type epithelium with goblet cells (endoderm) and cartilage (mesodermal), respectively. An arrow and arrowhead in the lower panel depict neuroecto-dermal (ectoderm) and muscle (mesoderm) tissues, respectively. Original magnification, 200. See also Figures S4 and S5 and Tables S4 and S5.

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Human embryonic stem cells derived by somatic cell nuclear ...