Liver-specific gene expression in mesenchymal stem cells is induced by liver cells.
OBJECTIVE
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.


CONCLUSIONS
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.
Lange
Claudia
C
Center of Bone Marrow Transplantation, University Hospital Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany. cllange@uke.uni-hamburg.de
Bassler
Philipp
P
Lioznov
Michael-V
MV
Bruns
Helge
H
Kluth
Dietrich
D
Zander
Axel-R
AR
Fiegel
Henning-C
HC
D000818:Animals; D015415:Biomarkers; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D008648:Mesoderm / Q000166:cytology; D051381:Rats; D011917:Rats, Inbred Lew
D015415:Biomarkers