人胚胎干细胞传代培养及细胞化学染色特性

目的 体外建立人胚胎干细胞传代培养方法,研究人胚胎干细胞细胞化学染色特性.方法 以小鼠胚胎成纤维细胞作为饲养层传代培养人胚胎干细胞,检测人胚胎干细胞、自发分化克隆及拟胚体的细胞化学染色特性.结果 人胚胎干细胞在小鼠胚胎成纤维细胞饲养层上传30代以上其形态保持不变；人胚胎十细胞碱性磷酸酶、过碘酸-雪夫反应、α-醋酸萘酚酯酶染色阳性,自发分化克隆细胞阳性程度明显减弱；人胚胎干细胞形成的拟胚体碱性磷酸酶染色弱阳性,过碘酸-雪夫反应、α-醋酸萘酚酯酶染色阳性.结论 小鼠胚胎成纤维细胞能支持人胚胎干细胞传代培养,细胞化学染色结果能初步鉴别人胚胎干细胞未分化特性.

Subculture and cytochemical staining characteristics of human embryonic stem cells

Objective To establish a method of subculture of human embryonic stem cells in vitro,and to study their cytochemical staining characteristics. Methods We subcultured human embryonic stem cells using mouse embryonic fibroblasts as the feeder layer,and explored the cytochemical staining characteristics of human embryonic stem cells,spontaneously differentiated colonies and embryoid bodies. Results Human embryonic stem cells whose morphology maintained consistancy were cultured more than 30 passages on mouse embryonic fibroblasts.The cytochemical staining of human embryonic stem cells for ALP,PAS and α-NAE were all positive,while the differentiated colonies became obviously weaker.In embryoid bodies,ALP was weak positive,whereas PAS and α-NAE were positive. Conclusion The mouse embryonic fibroblasts can support the subculture of human embryonic stem cells.Cytochemical staining can identify the characteristics of inundifferentiated human embryonic stem cells.