Study On Differentiation Of Rat Bone Marrow Stromal Cells Towards Liver Stem Cells In Vitro

Objective To explore the feasibility of differentiation of rat bone marrow stromal cells towards liver stem cells in vitro. Methods In vitro culture, we induced bone marrow stem cells to differentiate into hepatocyte-like cells by HGF(hepatocyte growth factor). Rat bone marrow stromal cells were treated with or without HGF on days 0, 7, 14, 21. The morphologic characteristics of the cells were observed; the alkaline phosphatase of cells were detected on day 7; The expression of AFP,albumin(ALB) and CK18 protein were detected immunohistochemically by using anti-AFP,albumin antibodies and CK18 antibodies; Glycogen were tested by periodic acid-schiff (PAS); The ultrastructure of the cells on day 21 with HGF induction and without induction were observed under inverted transmission electron microscope respectively. Results Under inverted microscope , the shape of cells with HGF induction changed from long Fusiform into round, lack of radiation disposition. Whereas in the groups without HGF induction, the cells are long Fusiform shape and array in radiation disposition or whirlpool. On day 7, the alkaline phosphatase of rat bone marrow stem cells were expressed strongly in HGF induction group,and AFP were expressed weakly in freshly isolated adult rat bone marrow stem cells , but didn't express ALB , CK18, and produce glycogen. In the HGF treated group, the AFP was positive at all time phase, however it tended to decrease after day 7; the expression of ALB, CK18 protein and glycogen–positive cells increased on the time–dependent manner.In the group without HGF,the AFP were detected weakly on days 7,but it could not be detected after then; ALB, CK18, glycogen could not be detected in every time phases. Under transmission electron microscope, the disposition of cells with HGF induction, many microvilli in the space between cells were observed, and...