摘要
目的探讨PDMSCs向肝细胞增殖和分化的体外培养条件及方法。方法孕20 d的大鼠无菌条件下取胎盘,经胶原酶消化、密度离心、贴壁筛选法分离培养胎盘源间充质干细胞,并对其表面抗原进行鉴定。在体外培养体系中加入胎肝滤液,模拟体内肝脏微环境,诱导PDMSCs向肝细胞定向分化,以免疫细胞化学检测干细胞标志物;PAS检测糖原表达。结果在体外培养条件下,PDMSCs贴壁生长为成纤维样细胞,CD44表面标志物检测阳性;PDMSCs经胎肝滤液诱导14d时细胞呈现圆形、卵圆形的特征性改变,AFP、CK19表达阳性。结论胎肝滤液能够诱导PDMSCs定向分化为肝细胞样细胞。
Objective To observe the differentiation of placental mesenchymal stem cells（PDMSCs） from E20d pregnant rats into hepatocytes induced by fetal liver filtrate,and explore the proliferation and differentiation conditions and induction methods of PDMSCs into hepatocytes cultured in vitro.Methods Placenta tissue was taken from the E20d pregnant rats under aseptic conditions.The placental tissue was digested by collagenase and centrifugation.The placenta-derived mesenchymal stem cells were isolated by adherence screening method and their surface antigen was identified.Fetal liver filtrate was added into the culture to simulate the environment in vivo and directionally induce PDMSCs to differentiate into hepatocytes.Immunocytochemistry was used to test the cellular markers,and PAS was used to detect glycogen expression.Results Under in vitro culture conditions,the PDMSCs developed into fibroblast-like cells,showing positive expression of CD44 surface marker.The cells appeared characteristically round and oval at 14 d when the PDMSCs were induced by fetal liver filtrate,and showing positive expressions of AFP and CK19.Conclusion Fetal liver filtrate can induce differentiation of PDMSCs into hepatocyte-like cells.
出处
《中国实验动物学报》
CAS
CSCD
2011年第5期419-422,I0006,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
山东省"泰山学者"建设工程专项经费
关键词
胎肝滤液
胎盘间充质干细胞
诱导分化
大鼠
肝细胞
Fetal liver filtrate
Placenta-derived mesenchymal stem cells
Differentiation
Rat
Hepatocytes
