Hepatocyte-like cells from directed differentiation of mouse bone marrow cells in vitro.
OBJECTIVE
To design the effective directed differentiation medium to differentiate bone marrow cells into hepatocyte-like cells.


METHODS
Bone marrow cells were cultured in the directed differentiation media including fibroblast growth factor-4 (FGF-4) and oncostatin M (OSM). Hepatocyte-like cells from directed differentiation of bone marrow cells were identified through cell morphology, RNA expressions by reverse transcriptase-polymerase chain reaction (RT-PCR), protein expressions by Western blot, and hepatocellular synthesis and metabolism functions by albumin ELISA, Periodic acid-Shiff staining and urea assay.


RESULTS
Some epithelial-like cells or polygonal cells appeared and increased in the course of the cell directed differentiation. Hepatocyte nucleur factor-3beta (HNF-3beta, albumin (ALB), cytokeratin 18 (CK18), transthyretin (TTR), glucose-6-phosphate (G-6-Pase), and tyrosine aminotransferase (TAT) mRNA were expressed in the course of the directed differentiation. The directed differentiated cells on d 21 expressed HNF-3? ALB, and CK18 proteins. The directed differentiated cells produced albumin and synthesized urea in a time-dependent manner. They could also synthesize glycogen.


CONCLUSIONS
Our differentiation media, including FGF-4 and OSM, are effective to differentiate bone marrow cells into hepatocyte-like cells, which could be used for hepatocyte resources for bioartificial liver or hepatocyte transplantation.
Shi
Xiao-lei
XL
Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, Medical College of Nanjing University, Nanjing 210008, China.
Qiu
Yu-dong
YD
Li
Qiang
Q
Xie
Ting
T
Zhu
Zhang-hua
ZH
Chen
Lei-lei
LL
Li
Lei
L
Ding
Yi-tao
YT
D000418:Albumins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D051519:Fibroblast Growth Factor 4; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D019298:Glucose-6-Phosphate / Q000096:biosynthesis / Q000235:genetics; D051542:Hepatocyte Nuclear Factor 3-beta; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D007633:Keratins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D053683:Oncostatin M; D010455:Peptides / Q000494:pharmacology; D011228:Prealbumin / Q000096:biosynthesis / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
D000418:Albumins; D004268:DNA-Binding Proteins; C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; C496503:Foxa2 protein, mouse; D009687:Nuclear Proteins; C508352:Osm protein, mouse; D010455:Peptides; D011228:Prealbumin; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D053683:Oncostatin M; D051542:Hepatocyte Nuclear Factor 3-beta; D019298:Glucose-6-Phosphate; D005346:Fibroblast Growth Factors; D007633:Keratins