Establishment Of In Vitro Hepatic Precursor Cells Bipotential Differentiation Models And Proteome Analysis Of WB-F344 Cells Differentiation Into Biliary Lineage Base On ITRAQ Technology

In the current study we differentiate the culture oval cells dissociated from 2-AAF/PH rat model and WB-F344 cell lines in vitro into the hepatocyte or cholangiocyte lineages selectively upon exposure the cells to sodium butyrate,DMSO, Matrigel and HGF/EGF. Immunocytochemical and RT-PCR results show that increased expressions of hepatocyte or cholangiocyte phenotypic markers and decrease of series stem cell phenotypic markers were confirmed in treated cells. Morphological changes were coincidence with the changes at the protein and mRNA levels. These results show that both rat oval cells and WB-F344 cells are able to differentiate bipotentially in vitro, making them candidate models for analyzing the molecular events associated with thedifferentiation of hepatic precusor cells. We applied the isobaric tags for relative and absolutequantitation on the proteomic study of WB-F344 cells during its in vitro differentiation along the biliary pathway. Proteome alterations were observed using elctrospray ionization tadem mass spectrometry, and alterationsin the proteome were examined. 1278 protein were found among those 67 proteins significantly altered. This study indicated that iTRAQ is a useful technologic method in proteomic alterations study of cell differentiation. The proteins revealed in this experiment can be used in the future for studies pertaining to hepatic precursor cells differentiation or as phenotypic marker for isolation and identification of hepatic stem cells.