ISOLATION OF A PRIMATE EMBRYONIC STEM-CELL LINE
ISOLATION OF A PRIMATE EMBRYONIC STEM-CELL LINE
复制标题
DOI：
10.1073/pnas.92.17.7844
复制
发表时间：
1995-08-15
影响因子：
11.1
通讯作者：
HEARN, JP
中科院分区：
文献类型：
--
作者：
THOMSON, JA;KALISHMAN, J;HEARN, JP
胚胎干细胞具有保持未分化和在体外无限增殖的能力，同时保持分化为所有三个胚胎胚层的衍生物的潜力。在这里，我们报告的衍生克隆细胞系（R278.5），其来自在连续传代中保持未分化>1年、维持正常XY核型并表达细胞表面标志物的恒河猴囊胚（碱性磷酸酶，阶段特异性胚胎抗原3，阶段特异性胚胎抗原4，TRA-1-60和TRA-1-（81）是人胚胎癌细胞的特征。当在小鼠胚胎成纤维细胞饲养层上生长时，R278.5细胞保持未分化，但在不存在成纤维细胞的情况下分化或死亡，尽管存在重组人白血病抑制因子。允许体外分化的R278.5细胞向培养基中分泌生物活性绒毛膜促性腺激素，表达绒毛膜促性腺激素α-和β-亚基mRNA，并表达甲胎蛋白mRNA，表明滋养层和内胚层分化。当注射到严重的联合免疫缺陷小鼠中时，R278.5细胞始终分化为所有三个胚胎胚层的衍生物。这些结果将R278.5细胞定义为胚胎干细胞系，据我们所知，这是第一个来自任何灵长类物种的细胞系。
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.