胚胎造血干Sca-1^＋细胞向肝细胞分化的研究.pdfVIP

第25卷第6期 广 东 医 学 院 学 报 Vo1．25 No．6 2007年12月 JOURNAL OF GUANGDONG MEDICAL COLLEGE Dec．2007 605 胚胎造血干Sca．1+细胞向肝细胞分化的研究 缪辉来 ，张晟晃 ，陈念平 ，陈 明 ，郑淑华 (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) 摘 要：目的 探讨胚胎造血干细胞抗原阳性(Sca—l )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—l 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 6 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-l 细胞能达到(85．57±1．66)％的纯度(P0．01)，对分离前后细胞活力的影响小 (P0．05)。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。 关键词：Sca—l 细胞；肝细胞；免疫磁性分离法；鉴定；分化 中图分类号：R 329．21 文献标识码：A 文章编号：1005 4057(2007)06—0605—05 Study on the differentiation from embryo haemopoietic cells positive of Sca-1 antigen to hepatic cells in vitro MIAO Hui—lai，ZHANG Sheng—huang，CHEN Nian—ping，CHEN Ming，ZHENG Shu—hua(Department of Hepatobiliary Surgery of the Affiliatted Hospital of Guangdong Medical College，Hepatobiliary Surgery Research Institute of Guangdong Medical College， Zha~iang 524001，China) Abstract：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 phse 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 immunohistochemical method．Result The purity Of Sca一1 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一1 cells transformed to hepatic cells in