Basic Research
Copyright ©The Author(s) 2005. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 7, 2005; 11(29): 4497-4504
Published online Aug 7, 2005. doi: 10.3748/wjg.v11.i29.4497
Liver-specific gene expression in mesenchymal stem cells is induced by liver cells
Claudia Lange, Philipp Bassler, Michael V. Lioznov, Helge Bruns, Dietrich Kluth, Axel R. Zander, Henning C. Fiegel
Claudia Lange, Philipp Bassler, Michael V. Lioznov, Axel R. Zander, Center of Bone Marrow Transplantation, University Hospital Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany
Helge Bruns, Dietrich Kluth, Henning C. Fiegel, Department of Pediatric Surgery, University Hospital Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany
Author contributions: All authors contributed equally to the work.
Supported by the “Rudolf Bartling Foundation” and “Foerderge-meinschaft Kinder-Krebs-Zentrum Hamburg e.V.”
Correspondence to: Dr. Claudia Lange, Center of Bone Marrow Transplantation, Universit鋞sklinikum Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany. cllange@uke.uni-hamburg.de
Telephone: +49-40-42803-5917 Fax: +49-40-42803-3795
Received: November 23, 2004
Revised: December 15, 2004
Accepted: December 20, 2004
Published online: August 7, 2005
Abstract

AIM: The origin of putative liver cells from distinct bone marrow stem cells, e.g. hematopoietic stem cells or multipotent adult progenitor cells was found in recent in vitro studies. Cell culture experiments revealed a key role of growth factors for the induction of liver-specific genes in stem cell cultures. We investigated the potential of rat mesenchymal stem cells (MSC) from bone marrow to differentiate into hepatocytic cells in vitro. Furthermore, we assessed the influence of cocultured liver cells on induction of liver-specific gene expression.

METHODS: Mesenchymal stem cells were marked with green fluorescent protein (GFP) by retroviral gene transduction. Clonal marked MSC were either cultured under liver stimulating conditions using fibronectin-coated culture dishes and medium supplemented with SCF, HGF, EGF, and FGF-4 alone, or in presence of freshly isolated rat liver cells. Cells in cocultures were harvested and GFP+ or GFP- cells were separated using fluorescence activated cell sorting. RT-PCR analysis for the stem cell marker Thy1 and the hepatocytic markers CK-18, albumin, CK-19, and AFP was performed in the different cell populations.

RESULTS: Under the specified culture conditions, rat MSC cocultured with liver cells expressed albumin-, CK-18, CK-19, and AFP-RNA over 3 weeks, whereas MSC cultured alone did not show liver specific gene expression.

CONCLUSION: The results indicate that (1) rat MSC from bone marrow can differentiate towards hepatocytic lineage in vitro, and (2) that the microenvironment plays a decisive role for the induction of hepatic differentiation of rMSC.

Keywords: Mesenchymal stem cells, Liver-specific differentiation, Coculture