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To investigate the possibility of multipotent adult progenitor cells from human bone marrow ZHJ-MAPC to differentiate into hepatocytes by co-culture with human hepatocyte line L02 in vitro.(1) Multipotent adult progenitor cells from human bone marrow, screened in Zhujiang hospital, named ZHJ-MAPC, were divided into 2 parts. Some ZHJ-MAPCs undergo co-culture with human hepatocytes of the line L-02 in the manner without cell to cell contact. On the days 1, 3, 5, and 7 immunocytochemistry was used to detect the expression of albumin (ALP), alpha-fetoprotein (AFP), cytokeratin (CK)-18, and CK-19, characteristic of hepatocyte. Some ZHJ-MAPC were labeled with Certified Functional Safety Expert (CFSE), a fluorescent reagent, and were mixed with L02 cells so as to undergo co-culture in the manner with cell to cell contact. Then the mixed cells were seeded on specialized dish for detection by Laser Scanning Confocal Microscope (LSCM). 5 days later, the cells were double stained with SABC-Cy3. The expression of ALP, AFP, and CK-18 in the ZHJ-MAPC were observed under LSCM. Separately cultured L02 hepatocytes served as positive control s and separately cultured ZHJ-MAPC served as negative controls.(1) Results of co-culture without cell to cell contact. On the first day, the ZHJ-MAPC expressed high level of AFP and then the AFP expression tapered daily. The expression of ALP was very weak on day 1 and then increased and peaked on day 5. The expression of CK-18 began to appear on day 5 and peaked on day 7. The expression of CK-19 was always negative. (2) Results of co-culture with cell to cell contact. On day 5 there were three colors of fluorescence under LSCM. Yellow cells represented the ZHJ-MAPC differentiating into hepatocytes; the green cells were undifferentiating ZHJ-MAPC; and the red cells were L02 hepatocytes. The result showed that ALP and CK-18 were expressed in many ZHJ-MAPC; AFP appeared only in a few cells.ZHJ-MAPC can be induced to differentiate into mature hepatocyte-like cells by co-culture, either with or without cell to cell contact. |
