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ISOLATION OF A PRIMATE EMBRYONIC STEM CELL LINE

机译:原始胚胎干细胞系的分离

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Embryonic stem cells have the ability to remain undifferentiated and proliferate indefinitely in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. Here we report the derivation of a cloned cell line (R278.5) from a rhesus monkey blastocyst that remains undifferentiated in continuous passage for >1 year, maintains a normal XY karyotype, and expresses the cell surface markers (alkaline phosphatase, stage-specific embryonic antigen 3, stage-specific embryonic antigen 4, TRA-1-60 and TRA-1-(81) that are characteristic of human embryonal carcinoma cells. R278.5 cells remain undifferentiated when grown on mouse embryonic fibroblast feeder layers but differentiate or die in the absence of fibroblasts, despite the presence of recombinant human leukemia inhibitory factor. R278.5 cells allowed to differentiate in vitro secrete bioactive chorionic gonadotropin into the medium, express chorionic gonadotropin alpha- and beta-subunit mRNAs, and express alpha-fetoprotein mRNA, indicating trophoblast and endoderm differentiation. When injected into severe combined immunodeficient mice, R278.5 cells consistently differentiate into derivatives of all three embryonic germ layers. These results define R278.5 cells as an embryonic stem cell line, to our knowledge, the first to be derived from any primate species. [References: 38]
机译:胚胎干细胞具有在体外保持未分化和无限增殖的能力,同时保持了分化为所有三个胚芽层的衍生物的潜力。在这里,我们报道了从恒河猴胚泡中克隆出的细胞系(R278.5)的衍生结果,该胚泡在连续传代> 1年的过程中保持未分化,保持了正常的XY核型,并表达了细胞表面标志物(碱性磷酸酶,阶段特异性胚胎抗原3,人类胚胎癌细胞特有的阶段特异性胚胎抗原4,TRA-1-60和TRA-1-(81)。R278.5细胞在小鼠胚胎成纤维细胞饲养层上生长时仍未分化,但可以分化或尽管存在重组人白血病抑制因子,R278.5细胞仍可在体外分化出分泌生物活性绒毛膜促性腺激素到培养基中,表达绒毛膜促性腺激素的α-和β-亚基mRNA,并表达α-甲胎蛋白。 mRNA,表明滋养层和内胚层的分化当注射入严重的免疫缺陷小鼠体内时,R278.5细胞持续分化为所有三个胚芽层。这些结果将R278.5细胞定义为胚胎干细胞系,据我们所知,这是第一个衍生自任何灵长类动物的细胞。 [参考:38]

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