[Study of mouse marrow cells differentiation into a hepatocyte lineage in vitro].

OBJECTIVE

To explore whether bone marrow stem cells (MSCs) from adult mice can be induced to differentiate into hepatocytes by hepatocyte growth factor (HGF) alone and the time phase characteristics in the differentiation progress.

METHODS

Adult mouse MSCs were treated with or without 100 ng/ml HGF, on days 0, 7, 14, 21, and 28. The morphologic characteristics of the cells were examined; the albumin (ALB), AFP mRNA was analyzed sub-quantively using reverse transcription polymerase chain reaction (RT-PCR) and immumohistochemistry techniques. The expression of ALB, AFP and CK19 were detected by using anti-ALB, AFP and CK19 antibodies.

RESULTS

Freshly isolated adult mouse MSCs expressed ALB and AFP mRNA weakly; in the group without HGF, no ALB mRNA was detected on day 7. The expression of AFP mRNA was reduced significantly on day 7, and could not be detected anymore after day 14. In the HGF treated group, ALB mRNA was not detected on day 7, but the positive lane appeared again on day 14, and the expression of ALB mRNA was increased on day 21 but reduced in the following days. The AFP mRNA was positive at all times, however it tended to decrease after day 14 in the HGF treated groups. The result of immumohistochemistry was consistent with that of RT-PCR, and CK19 was always negative.

CONCLUSION

Adult mouse MSCs can be induced into hepatocyte differentiation in vitro. The optimal time for the induction was 2 to 3 weeks.