Differentiation of rat bone marrow cells cultured on artificial basement membrane containing extracellular matrix into a liver cell lineage.
OBJECTIVE
Bone marrow (BM) cells have been shown to be capable of differentiating into a liver cell lineage in vitro. However, their differentiation and proliferation is poor, and the cell characteristics are poorly understood.


METHODS
We cultured rat BM cells on an artificial basement membrane containing extracellular matrix (ECM) with hepatocyte growth factor (HGF). The expression of mRNA for liver-specific genes was analyzed by reverse transcription PCR. The expression of albumin and Musashi-1 by cultured cells was analyzed using a fluorescence-activated cell sorter (FACS). The proportions of albumin-positive cells when culture was performed with different concentrations of HGF were analyzed by FACS.


RESULTS
On culture day 21, polygonal cells proliferated and formed cell colonies. These cells expressed mRNA for all the liver-specific genes analyzed, and showed heterogeneous differentiation, some cells expressing albumin, others expressing Musashi-1. Albumin-positive differentiated cells were large and rich in intracellular structures, while Musashi-1-positive undifferentiated cells had the opposite characteristics. Culturing cells with higher concentrations of HGF induced an increased proportion of albumin-positive cells.


CONCLUSIONS
The results suggest that cell culture on an ECM with a high concentration of HGF increases the extent to which BM cells differentiate into a liver cell lineage and proliferate in vitro.
Okumoto
Kazuo
K
Division of Gastroenterology, Department of Internal Medicine and Molecular Therapeutics, Yamagata University School of Medicine, Yamagata 990-9585, Japan.
Saito
Takafumi
T
Hattori
Etsuko
E
Ito
Jun-itsu
JI
Suzuki
Akihiko
A
Misawa
Keiko
K
Ishii
Rika
R
Karasawa
Tetsuru
T
Haga
Hiroaki
H
Sanjo
Mai
M
Takeda
Tadashi
T
Sugahara
Kazuhiko
K
Saito
Koji
K
Togashi
Hitoshi
H
Kawata
Sumio
S
D000418:Albumins / Q000378:metabolism; D000818:Animals; D001485:Basement Membrane; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division; D019070:Cell Lineage; D002469:Cell Separation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000502:physiology*; D005434:Flow Cytometry; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000008:administration & dosage / Q000494:pharmacology; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D008297:Male; D008567:Membranes, Artificial; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009994:Osmolar Concentration; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D000418:Albumins; D015415:Biomarkers; D008567:Membranes, Artificial; C491296:Msi1 protein, rat; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D017228:Hepatocyte Growth Factor