人胚胎肝细胞的体外培养及表型观察

基础医学与临床 ›› 2007, Vol. 27 ›› Issue (4): 386-390.

人胚胎肝细胞的体外培养及表型观察

杨帆王春景苏晓玲

医科院基础医学研究所医科院基础医学研究所医科院基础医学研究所

收稿日期:2005-10-31 修回日期:2006-10-17 出版日期:2007-04-25 发布日期:2007-04-25
通讯作者: 杨帆

The culture and phenotypic examination of human fetal hepatocytes in vitro

Received:2005-10-31 Revised:2006-10-17 Online:2007-04-25 Published:2007-04-25

摘要/Abstract

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种特异性酶的生物功能。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。

Abstract: 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 quantity 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 and keep function in vitro for several months.

杨帆王春景苏晓玲. 人胚胎肝细胞的体外培养及表型观察[J]. 基础医学与临床, 2007, 27(4): 386-390.

. The culture and phenotypic examination of human fetal hepatocytes in vitro[J]. Basic & Clinical Medicine, 2007, 27(4): 386-390.