Hepatocyte-like cells from directed differentiation of mouse bone marrow cells in vitro
Abstract
Aim: 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-3β (HNF-3β), 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-3β, ALB, and CK18 proteins. The directed differentiated cells produced
albumin and synthesized urea in a time-dependent manner. They could also synthesize
glycogen. Conclusion: 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.
Keywords:
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-3β (HNF-3β), 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-3β, ALB, and CK18 proteins. The directed differentiated cells produced
albumin and synthesized urea in a time-dependent manner. They could also synthesize
glycogen. Conclusion: 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.