Purification of adult hepatic progenitor cells using green fluorescent protein (GFP)-transgenic mice and fluorescence-activated cell sorting

被引:41
作者
Fujikawa, T
Hirose, T
Fujii, H
Oe, S
Yasuchika, K
Azuma, H
Yamaoka, Y
机构
[1] Kyoto Univ, Grad Sch Med, Dept Surg Gastroenterol, Sakyo Ku, Kyoto 6068507, Japan
[2] Kyoto Univ, Inst Frontier Med Sci, Dept Regenerat Med, Sakyo Ku, Kyoto 6068507, Japan
基金
日本学术振兴会;
关键词
adult hepatic progenitor cell; green fluorescent protein; transgenic mouse; fluorescence-activated cell sorting; hepatocyte; biliary epithelial cell; regenerative medicine;
D O I
10.1016/S0168-8278(03)00237-X
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Background/Aims: Recent advances in stem cell research have revealed that hepatic stem/progenitor cells may play an important role in liver development and regeneration. However, a lack of detectable definitive markers in viable cells has hindered their primary culture from adult livers. Methods: Enzymatically dissociated liver cells from green fluorescent protein (GFP)-transgenic mice, which express GFP highly in liver endodermal cells, were sorted by GFP expression using a fluorescence-activated cell sorter. Sorted cells were characterized, and also low-density cultured for extended periods to determine their proliferation and clonal differentiation capacities. Results: When CD45-TER119(-) side-scatter(low) GFP(high) cells were sorted, alpha-fetoprotein-positive immature endoderm-characterized cells, having high growth potential, were present in this population. Clonal analysis and electron microscopic evaluation revealed that each single cell of this population could differentiate not only into hepatocytes, but also into biliary epithelial cells, showing their bilineage differentiation activity. When surface markers were analyzed, they were positive for Integrin-alpha6 and -beta1, but negative for c-Kit and Thy1.1. Conclusions: Combination of GFP-transgenic mice and fluorescence-activated cell sorting enabled purification of hepatic progenitor cells from adult mouse liver. Further analysis of this population may lead to purification of their human correspondence that would be an ideal cell-source candidate for regenerative medicine. (C) 2003 Published by Elsevier Science B.V. on behalf of European Association for the Study of the Liver.
引用
收藏
页码:162 / 170
页数:9
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