胚胎造血干Sca-1^+细胞向肝细胞分化的研究  

Study on the differentiation from embryo haemopoietic cells positive of Sca-1^+ antigen to hepatic cells in vitro

作　　者：缪辉来[1] 张晟晃[1] 陈念平[1] 陈明[1] 郑淑华[2] 

机构地区：[1]广东医学院附属医院肝胆外科 [2]广东医学院肝胆外科研究室,广东湛江524001

出　　处：《广东医学院学报》2007年第6期605-608,613,共5页Journal of Guangdong Medical College

基　　金：广东省社会发展领域科技计划项目(20053026);湛江市科技招标项目基金(200515)

摘　　要：目的探讨胚胎造血干细胞抗原阳性(Sca-1^+)细胞亚群在合适的体外诱导体系下向肝细胞或肝细胞前体分化的可行性。方法免疫磁性分离法分离Sca-1^+细胞,相差显微镜观察细胞形态,RT-PCR法检测细胞白蛋白(ALB)、转甲状腺蛋白mRNA水平表达,免疫组织化学法检测AFP和CK8/18蛋白水平的表达,以及检测细胞糖原染色和尿素合成功能。结果免疫磁性分离法分离Sca-1^+细胞能达到(85.57±1.66)%的纯度(P<0.01),对分离前后细胞活力的影响小(P<0.05)。随着诱导分化时间延长,细胞形态明显向肝细胞变化,AFP表达消失、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。结论免疫磁性分离法能够分离出较高纯度的胚胎造血干Sca-1^+细胞,并能够向肝脏细胞分化。Objective To investigate the differentiation from embryo haemopoietic cells positive of Sca-1^＋ antigen to hepatic celsl or hepatic cell precursors under a suitable induction in vitro. Methods Sca-1^＋ cells were isolated by immunization magnetic separation and their morphology was observed with contrast phase microscope. The mRNA expressions of albumin （ALB） and transthyroprotein genes were detected by RT-PCR. The AFP and CK8/18 protein, cell glycogen and urea synthesis were detected by immunohistochemlcal method. Result The purity of Sca-1^＋ cells separated by immunization magnetic separation reached to （85.57 ＋ 1.66） without affect ting the activity of the cells. With the induction going, the Sca-1^＋ cells transformed to hepatic cdls in morphology with down expression of AFP and up expression of CK8/18 protein, and mRNA of ALB and transthyroprotein genes. When the Sca-1^＋ cells differentiated to mature hepatic cells, the cell glycogen and urea synthesis function were strengthened. Conclusion The pure Sca-1^＋ cells could be isolated from embryo haemopoietic cells with immunization magnetic separation and induced to differentiate to mature hepatic cells in vitro.

关 键 词：SCA-1^+细胞 肝细胞 免疫磁性分离法 鉴定 分化 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]