人胚胎干细胞饲养层培养体系的建立  

ESTABLISHMENT OF CULTURE SYSTEM ON HUMAN EMBRYONIC STEM-LIKE CELLS FEED LAYER

作　　者：苗增民[1] 李松[2] 窦忠英[3] 

机构地区：[1]山东农业大学 [2]泰山医学院 [3]西北农林科技大学

出　　处：《济宁医学院学报》2005年第3期4-5,共2页Journal of Jining Medical University

基　　金：国家自然科学基金(39970363);国家973项目资助(G1999054300)

摘　　要：目的建立人胚胎干细胞饲养层培养体系。方法以胎鼠和人胎儿为材料,以DMEM加15%FBS加0.1mM2ME为基础培养液,分别制备胎鼠和胎儿成纤维细胞饲养层。结果分别获得了传16代的胎鼠成纤维细胞和传7代的人胎儿成纤维细胞,建立了人胚胎干细胞饲养层的培养体系。结论胎鼠成纤维细胞和人胎儿成纤维细胞相比较,后者生长速度慢,且较易老化;胎鼠成纤维细胞饲养层更有利于人胚胎干细胞的生长;8～14d龄胎鼠最适宜制备人胚胎干细胞饲养层;在室温条件下,以0.2%胰酶加0.04%EDTA为消化液处理原代胎鼠组织块时间以15～20min为宜。Objective To establish the culture system of human embryonic stem- like cells feed layer. Methods The culture system were composed of DMEM supplemented with 15 % NBS,0.1 mM beta- mercaptoethanol, we prepared the feed lay about fetus mouse fibroblast cells and human fetus fibroblast. Results Acquired 16 passages in mouse and 7 passages in human and established the culture system of human embryonic stein-like ceils. Conclusion Human fetus fibroblast grew lower and easily aging than mouse fibroblast, the former was better for growing of human embryonic stem- like cells;mouse fetus of 8-14 days old was proper to prepare feed layer; In general,it was proper that mouse fetus＇ tissue was dealed for 15-20min in the condition of 0.2 % trypsin and 0.04 % EDTA.

关 键 词：人胚胎干细胞 饲养层 成纤维细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R321-33[医药卫生—基础医学]