Rat mesenchymal stem cells differentiate into hepatocyte-like cells induced hepatocyte growth factor plus epidermal growth factor in vitro / 解放军医学杂志

ABSTRACT

Objective To research the possibility of the rat mesenchymal stem cells (MSCs) to differentiate into hepatocyte-like with hepatocyte growth factor plus E growth factor in vitro. Methods (1) Bone marrow in the femurs of Wistar rat was collected by flushing under sterile condition. MSCs were separated and cultured according to the direct anchoring method, identified by using immunocytochemical methods and flow cytometry. After MSCs were induced by HGF (20mg/ml) and EGF (10mg/ml), mRNA expression level of AFP、Alb、CK-18 in the MSCs were detected by RT-PCR. In addition, ultrastructures in the cells induced were observed using electron microscope. Results Anchored MSCs is single or cell clone is developed. Cell is uniform in the configuration and converged long fusiform after 7-10 days. Cultured cells are certified to be MSCs in the immunocytochemistry and flow cytometry. Afer 21 days, cultured cells show Hepatocyte-like characters in the configuration. RT-PCR:On day 7 mRNA of AFP was detected; mRNA of Alb and CK-18 were detected at the beginning, then strengthen on 21 day. At the same time, induced cells in the electron microscope results coincided with hepatic cells. Conclusions MSCs can be isolated, cultured and expanded easily in vitro. MSCs induced by HGF plus EGF in vitro can differentiate into hepatocyte-like cells. As a result, it can provide a new therapeutic method for liver failure.