[Proliferation and differentiation of C-kit+ cell in vitro]

This study sought to isolate and purify C-kit+ cells from rat 2-AAF/PH model and to investigate the proliferation and differentiation of C-kit+ cells in vitro. C-kit positive oval cells were enriched by using magnetic activated cell sorting (MACS). The sorted oval cells were cultured in a low density, and then colony formation was observed. The capacity of proliferation and differentiation of C-kit positive cells were examined in vitro by immunocytochemistry and RT-PCR. By using C-kit antibody in conjunction with MACS, we developed a rapid oval cell isolation protocol. The sorted cells formed colony when cultured in vitro. Cells in the colony expressed albumin or cytokeratin 19 (CK19) or coexpressed both, and BrdU incorporation test was positive. RT-PCR on colony showed expression of albumin and CK19 gene. The results demonstrate that by means of MACS we have established a method to isolate oval cells. The sorted hepatic oval cells can form colony in vitro which expresses different combinations of phenotypic markers and genes from both hepatocytes and cholangiocyte lineage.