J Korean Soc Transplant.
2006 Jun;20(1):35-40.
In vitro Differentiation of Human Mesenchymal Stem Cells into Hepatocytes
- Affiliations
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- 1Stem Cell Therapy Center, Soonchunhyang University Hospital, Korea. dhchoi@hosp.sch.ac.kr
- 2Department of Surgery, College of Medicine, Soonchunhyang University, Seoul, Korea.
- 3Department of Internal Medicine, College of Medicine, Soonchunhyang University, Seoul, Korea.
- 4Department of Radiology, College of Medicine, Soonchunhyang University, Seoul, Korea.
- 5Department of Urology, College of Medicine, Soonchunhyang University, Seoul, Korea.
Abstract
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PURPOSE: Whole liver transplantation has limitation including donor shortage and fatal surgical complications. Hepatocyte transplantation, which is simpler and less expensive than whole liver transplantation, allows the use of living related donors, permits the use of a single donor organ for multiple recipients, and makes possible the cryopreser-vation of hepatocytes for future use. However, hepatocytes have limitation of proliferation and lose their property during culture period. To over come this problems, here we performed differentiation of bone marrow derived mesenchymal stem cells into hepatocytes.
METHODS
Human bone marrow cells were harvested from posterior iliac spine of male and then mononuclear cells were obtained by Ficoll-Paque density-gradient centrifuge and plated in tissue culture flasks. For hepatogenic differentiation, we used modified Kuan-Der Lee's method. After differentiation, hepatocytes were collected and RT-PCR and PAS stain analysis were performed.
RESULTS
After 5 weeks of cultivation period, mesenchymal stem cells showed cuboidal morphology and contained abundant granules in the cytoplasm. RT-PCR analysis showed increased expression of hepatocyte-specific marker genes (albumin,CK18, PERCK, CPS). Undifferentiated MSCs were not stained with PAS and differentiated hepatocytes from human MSCs stained with PAS indicating that hepatocytes contained glycogen in the cytoplasm.
CONCLUSION
Hepatocyte transplantation could be one of the most effective treatments for chronic liver disease. However, hepatocyte has several disadvantages and problems. For alternative cell therapy sources, human bone marrow derived MSCs are considered as transplantable cells. Human MSCs are able to differentiate into functional hepatocytes in vitro and can be a possible cell transplantation source for chronic liver disease patients. Further studies should be done for differentiating human MSCs to hepatocytes in vivo condition.