Wang Ping, Zhang Haiyan, Li Weihong, Zhao Yongmei, An Wei
Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing, 100069, China.
Histochem Cell Biol. 2008 Aug;130(2):375-85. doi: 10.1007/s00418-008-0439-2. Epub 2008 May 14.
Hepatoblasts, which are considered one type of hepatic progenitor cell, reside in the fetal liver. To selectively identify these cells, we transfected primary cultured human fetal liver cells (FLCs) with a pGL3 vector bearing the gene for the enhanced green fluorescence protein (EGFP) under the control of the alpha-fetoprotein (AFP) promoter expressed in hepatoblasts. The FLCs were then sorted by fluorescence-activated cell sorting (FACS) on the basis of AFP promoter-driven EGFP expression. The EGFP-positive cells expressed AFP, albumin, and cytokeratin 19, and could be expanded in vitro. Thus, the AFP promoter-EGFP reporter system is highly useful for identification and isolation of hepatic progenitor cells.
肝母细胞被认为是一种肝祖细胞，存在于胎儿肝脏中。为了选择性地识别这些细胞，我们用一个pGL3载体转染原代培养的人胎儿肝细胞（FLCs），该载体带有在肝母细胞中表达的甲胎蛋白（AFP）启动子控制下的增强型绿色荧光蛋白（EGFP）基因。然后根据AFP启动子驱动的EGFP表达，通过荧光激活细胞分选（FACS）对FLCs进行分选。EGFP阳性细胞表达AFP、白蛋白和细胞角蛋白19，并且可以在体外扩增。因此，AFP启动子-EGFP报告系统对于肝祖细胞的识别和分离非常有用。
