肝细胞生长因子与酸性成纤维细胞生长因子体外诱导胚胎肝干细胞抗原阳性细胞向肝细胞定向分化的研究

目的探讨肝细胞生长因子（HGF）与酸性成纤维细胞生长因子（aFGF）体外诱导胚胎肝干细胞抗原阳性（Sca-1^＋）细胞向肝细胞或肝细胞前体分化的可行性。方法免疫磁珠分离法分离Sca-1^＋细胞，相差显微镜观察细胞形态，（RT-PCR半定量逆转录聚合酶链反应）法检测细胞自蛋白（ALB）、转甲状腺蛋白mRNA水平表达；免疫组织化学法检测ALB、AFP和CKS／18蛋白水平的表达，以及检测细胞糖原染色和尿素合成功能。结果分离后Sca-1^＋细胞活力为（94．24±1．04）％，纯度为（85．57±1．66）％，回收率为（62．31±1．85）％。Sca-1^＋细胞经HGF和aFGF诱导分化后，细胞形态变成不规则，明显向肝细胞变化，AFP蛋白表达消失、ALB和CK8／18蛋白表达升高，ALB、转甲状腺蛋白mRNA表达升高，且细胞糖原染色和尿素合成功能增强，逐渐向成熟肝细胞转化。结论HGF与aFGF可在体外诱导胚胎肝Sca-1^＋细胞向肝细胞或肝细胞前体分化。

Study on the orient differentiation from embryo hepatic Sca-1+ cell to hepatic cell under the induction of HGF and aFGF in vitro

Objective To investigate the feasbility of the orient differentiation from embryo hepatic Sca-1+ cell to hepatic cell or hepatic cell precursor under the induction of hepatocyte growth factor(HGF) and acor fibroblast growth factor(aFGF) in vitro. Methods Sea-1+ cells were isolated by immunomagnetic beads and their morphous were observed by contrast phase microscope. The expression of mRNA of albumin(ALB) and meta-thyroprotein were detected by RT-PCR; ALB, AFP, CK8/18 protein were detected by immunohistochemical method, and cell staining for glycogen and urea synthesis function were tested. Results The activity, purity, recovery rate of Sca-1+ cells were (94. 24±1.04) %, (85.57±1.66) %, (62. 31±1.85 ) % respectively. After the induction of HGF and aFGF, Sca1+ cells became anomalism and transformed to heptic cell in morphous,disappeared in expression of AFP protein and upregulated in expression of ALB and CK8/18 protein,upregulated in expression of ALB and transthyroprotein mRNA,cell staining of glycogen and urea synthesis function were strengthened,the cell differentiated to mature hepatic cell. Conclusion Embryo hepatic Sca-1+ cell could differentiate to hepatic cell or hepatic cell precursor under the induction of HGF and aFGF in vitro.