Hepatocytic differentiation of mesenchymal stem cells in cocultures with fetal liver cells

被引:38
作者
Lange, Claudia
Bruns, Helge
Kluth, Dietrich
Zander, Axel R.
Fiegel, Henning C.
机构
[1] Univ Hamburg, Hosp Eppendorf, Ctr Bone Marrow Transplantat, D-20246 Hamburg, Germany
[2] Univ Hamburg, Hosp Eppendorf, Dept Pediat Surg, D-20246 Hamburg, Germany
关键词
hepatic stem cells; mesenchymal stem cells; fetal liver cells; co-culture;
D O I
10.3748/wjg.v12.i15.2394
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
AIM: To investigate the hepatocytic differentiation ofmesenchymal stem cells (MSCs) in co-cultures with fetal liver cells (FLC) and the possibility to expand differentiated hepatocytic cells. METHODS: MSCs were marked with green fluorescent protein (GFP) by retroviral gene transduction. Clonal marked MSCs were either cultured under liver stimulating conditions using fibronectin-coated culture dishes and medium supplemented with stem cell factor (SCF), hepatocyte growth factor (HGF), epidermal growth factor (EGF), and fibroblast growth factor 4 (FGF-4) alone, or in presence of freshly isolated FLC. Cells in co-cultures were harvested, and GFP+ or GFP- cells were separated using fluorescence activated cell sorting. Reverse transcription-polymerase chain reaction (RT-PCR) for the liver specific markers cytokeratin-18 (CK-18), albumin, and alpha-fetoprotein (AFP) was performed in different cell populations. RESULTS: Under the specified culture conditions, rat MSCs co-cultured with FLC expressed albumin, CK-18, and AFP-RNA over two weeks. At A 3, MSCs lost hepatocytic gene expression, probably due to overgrowth of the cocultured FLC. FLC also showed a stable liver specific gene expression in the co-cultures and a very high growth potential. CONCLUSION: The rat MSCs from bone marrow can differentiate hepatocytic cells in the presence of FLC in vitro and the presence of MSCs in co-cultures also provides a beneficial environment for expansion and differentiation of FLC. (c) 2006 The WIG Press. All rights reserved.
引用
收藏
页码:2394 / 2397
页数:4
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