Abstract:

BACKGROUND: Studies have confirmed that in a variety of micro-environments, bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into mature hepatocytes, but the inducible condition and differentiating rate are inconclusive. To select the appropriate inducer and its concentration is particularly important.
OBJECTIVE: Through study on the differentiation of rabbit BMSCs into hepatocytes in vitro induced by different concentrations of hepatocyte growth factor (HGF) and epidermal cell growth factor (EGF), to find the best concentration combination of HGF and EGF.
METHODS: BMSCs were induced with different concentrations of HGF and HGF. The morphological changes of cells were observed. In different culture periods, the hepatic surface phenotype including alpha fetoprotein (AFP), albumin (ALB) and synthesis function of hepatocytes were determined.
RESULTS AND CONCLUSION: After prolonged culture, hepatocyte-like cells were seen. Until day 7, AFP was positive, but reduced in the following days (P < 0.05), and after that, there were no difference among all groups (P > 0.05). Until day 14, ALB expression was positive, and increased with the culture time (P < 0.05). The higher the concentration of cell growth factors, the higher the number of positive cells (P< 0.05). The concentration of ALB in the supernatant in the early stage (9-15 days) showed a concentration-dependent manner, and reached the peak at 18 or 21 days. Then the concentration began to decreased, and there were no difference (P > 0.05). The high concentrations of HGF and EGF may enhance the rate of differentiation, and the best combination is HGF 60 ug/L and EGF 4.5 mg/L.