title: Hepatocytic differentiation of mesenchymal stem cells in cocultures with fetal liver cells.
abstract: OBJECTIVE
  To investigate the hepatocytic differentiation of mesenchymal 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 wk 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.
  
  
  CONCLUSIONS
  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.
authors: Center of Bone Marrow Transplantation, Universitatsklinikum Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany. cllange@uke.uni-hamburg.de
authors: Claudia
authors: C
authors: Lange
authors: Helge
authors: H
authors: Bruns
authors: Dietrich
authors: D
authors: Kluth
authors: Axel-R
authors: AR
authors: Zander
authors: Henning-C
authors: HC
authors: Fiegel