基础医学与临床 ›› 2009, Vol. 29 ›› Issue (5): 535-538.

• 技术与方法 • 上一篇    下一篇

诱导小鼠原始生殖细胞向肝细胞样细胞分化的新方法

赵化芳 李晓燕 史小林   

  1. 首都医科大学生殖医学研究中心 首都医科大学生殖医学研究中心
  • 收稿日期:2008-09-19 修回日期:2008-12-04 出版日期:2009-05-25 发布日期:2009-05-25
  • 通讯作者: 史小林

The new method of inducing mouse primordial germ cells into hepatocytes-like in vitro

Hua-fang ZHAO, Xiao-yan LI, Xiao-lin SHI   

  1. Center of Reproductive Medicine, Capital Medical University Center of Reproductive Medicine, Capital Medical University
  • Received:2008-09-19 Revised:2008-12-04 Online:2009-05-25 Published:2009-05-25
  • Contact: Xiao-lin SHI

摘要: 目的 探索诱导小鼠原始生殖细胞向肝细胞样细胞定向分化的有效方法。方法 分离孕13d昆明小鼠的原始生殖细胞(PGC),体外培养扩增后,向其培养液中加入新生鼠肝组织提取液和包被有胎肝细胞的微囊共培养,利用微环境诱导法使PGC定向分化,然后免疫细胞化学法标记α-1-抗胰蛋白酶(AAT)和白蛋白(ALB)。结果 在微环境的诱导作用下,可见PGC分化成卵圆形或星形肝细胞样细胞,并且分化细胞的数量随诱导时间延长而增多,培养2周时分化率可达70%以上。分化的肝细胞样细胞,AAT和ALB均呈阳性表达。结论 应用新生小鼠肝组织提取液和包被有胎肝细胞的微囊构成的微环境,可有效诱导PGC向肝细胞方向分化。

Abstract: Objective To establish a new method for inducing the primordial germ cells to differentiate into hepatocyte-like in vitro. Methods The primordial germ cells (PGCs) from the gonadal ridges of the mouse embryos at 13 days postcoitum from Kunming pregnant mice were cultured and proliferated in vitro. Then embryonic hepatocytes enclosed in microcapsule and liver tissue extract of newborn mice were added into medium to co-culture with PGCs for committed differentiation. Albumin (ALB) and α-1-antitrypsin (AAT) were assayed by immunocytochemistry. Results The morphology of cells differentiated from PGCs likes star or ovum, the ALB and AAT immune positive expression were detected in those differentiated cells. The ratio of positive cells was above 70% in 2 weeks. Conclusion Microenvironment of embryonic hepatocyte microcapsules and liver tissue extract could effectively induce PGCs to differentiate into hepatocytes.