小鼠胚胎干细胞体外分步诱导为肝细胞的实验研究

Differentiation of mouse embryonic stem cells into hepatocytes in vitro

AIM: To explore a method for the differentiation of embryonic stem (ES) cells into hepatocytes in vitro. METHODS: ES cells were cultured on the feeder layer of mouse embryonic fibroblasts (MEF) in the medium containing leukemia inhibitory factor (LIF). The inductive factors, such as acid-fibroblast growth factor (acid-FGF), HGF, oncostatin M (OSM), Dex and ITS mixture (5 g/L insulin, 5 g/L transferring, 5 mg/L selenic acid), were added at day 9,12,12,15 of induction. The activity of alkaline phosphatase (ALP) was detected by cytochemical method and visualized by NBT/BCIP. Albumin (ALB) and CK18 were examined by immunohistochemistry and DAB staining. RESULTS: ALP was positive in the undifferentiated ES cells. Four days after induction, embryonid bodies came into formation. After the inductive factors were added, ES cells were differentiated into single morphological cells like epithelium. ALB and CK18 were positive in some differentiated cells. CONCLUSION: Embryonic stem cells can be selectively differentiated into hepatocytes in vitro.