人类饲养层及无血清培养体系培养人类胚胎生殖细胞研究

使用人类胚胎皮肤成纤维细胞(hESF)作饲养层,以血清替代物及一些细胞因子作无血清培养基,培养人类胚胎生殖(hEG)细胞。结果显示,在该培养体系下,人类胚胎生殖细胞能够存活、增殖,并表现出干细胞特有的形态学特征,具有很高的碱性磷酸酶(AP)活性,在体外能够保持未分化状态50 d左右。接种hEG细胞后,该体系能够比以前的人类胚胎生殖细胞培养体系形成更多的集落,并且这些集落AP活性维持更久。结果为人类胚胎生殖细胞的基础研究和临床应用提供了一种更为安全的培养方法。

Human cmbryonic skin fibroblasts and serum-free culture for human embryonic germ cells

In this study,we proved that human embryonic skin fibroblasts(hESFs) feeder layers plus a serum-free culture medium,consist of Knock-out serum replacer(KSR) and some cell factors could support the survival and proliferation of the human embryonic germ(hEG) cells,which demonstrated normal morphological characteristics of stem cell,possessed positive alkaline phosphatase(AP) activity,and could be maintained undifferentiated state for about 50 days;More colonies were formed in this culture system which could possess AP activity for relatively longer period than the previous hEG culture system when the same amount of cells was plated.These results provide a safer alternative culture protocol for basic research and clinical applications of these cells.