人羊膜上皮细胞的干细胞特性

Stem cell characteristics of human amniotic epithelial cells

背景:研究发现人羊膜上皮细胞具有类似胚胎干细胞或多能干细胞的多向分化潜能,说明其可能是未来组织工程重建的一种新型种子细胞。目的:研究体外培养的人羊膜上皮细胞的干细胞特性。方法:取足月剖宫产的人羊膜组织,经酶消化法和差异黏附法获得纯度高的人羊膜上皮细胞,接种于含体积分数为10%胎牛血清的DMEM/F12培养基中进行原代和传代培养,用免疫荧光法和流式细胞仪检测法检测人羊膜上皮细胞表面胚胎干细胞的表面标记蛋白OCT-4和干细胞表面分子标记CD29、CD34、CD44、CD45、CD105的表达。结果与结论:人羊膜上皮细胞在体外培养条件下呈上皮细胞特有的铺路石样外观,其胞浆中有OCT-4免疫荧光表达,其干细胞标记分子CD29、CD34的表达是阳性,但干细胞标记分子CD44、CD45、CD105的表达为阴性。结果表明人羊膜上皮细胞具有干细胞的某些特性,其可能是未来组织工程重建的一种新型种子细胞。

BACKGROUND： Studies have found that human amniotic epithelial cells （hAECs） have a multiple differentiation capacity similar as pluripotent stem cells or embryonic stem cells.It is indicated that hAECs may be a new kinds of seed cells in tissue engineering reconstruction in the further.OBJECTIVE： To culture the hAECs in vitro and to investigate stem cell characteristics of cultured hAECs.METHODS： The human amniotic tissue was delivered from full term caesarean,and the highly purified hAECs were obtained from enzyme digestion method and differently sticky method.The hAECs were seeded in DMEM/F12 medium obtaining 10% fetal bovine serum for the first time and serial subcultivation.The immunofluorescence and flow cytometry were used to detect the expression of OCT-4 of stem cell factor and Stem cell surface molecule marker CD29,CD34,CD44,CD45,CD105 in passaged hAECs.RESULTS AND CONCLUSION： The cells in vitro looked like slabstone that epithelial cells specified.The results of immunofluorescence showed the expression of OCT-4 of stem cell factor in hAECs was positive.Detection of stem cell surface molecule marker by the flow cytometry showed the expression of CD29,CD34 on hAECs were positive,and the expression of CD44,CD45,CD105 on hAECs were negative.The findings show that the hAECs have some characteristics of embryonic stem cells,and may be a new kind of seed cells for the future tissue engineering reconstitution.