人胚胎肝细胞的体外培养及表型观察  

Culture and characterization of human fetal hepatocytes

作　　者：杨帆[1] 王春景[1] 苏小玲[1] 顾蓓[1] 张宏[1] 刘玉琴[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所,北京100005

出　　处：《基础医学与临床》2007年第4期386-390,共5页Basic and Clinical Medicine

基　　金：国家科技部基础性研究项目(2000-19)

摘　　要：目的观察体外培养的人胚胎肝细胞的特性。方法采用胰酶分步消化法体外分离人胚胎肝细胞,在培养的不同代数,收集培养上清液测定甲胎蛋白、白蛋白及5种特异性酶(ALT、AST、GGT、ALP、LDH)的分泌量,用免疫组化法测定细胞内细胞色素C的表达情况。观察诱导分化剂丁酸钠对该细胞的作用效果。结果体外分离培养的肝细胞在DMEM培养基中呈单层多边形上皮样生长,常规传代可维持约5个半月(30代),在传代过程中具有分泌白蛋白及5种特异性酶的生理功能。结论建立的人胚胎肝细胞系,保持了肝细胞的表型及分泌多种特异性酶。Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro ; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes （ including ALT, AST, GGT, ALP and LDH） of different generations in culture ; determining the expression of cytochrome C by immunohistochemistry ; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months （ about 30 passages） in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

关 键 词：细胞培养 肝细胞 胚胎 丁酸钠 

分 类 号：Q813.1[生物学—生物工程]