摘要
目的比较两种体外分离培养原代大鼠胚胎肝干细胞(EHSCs)方法,并对其进行鉴定。方法用胶原酶灌注法及剪碎消化法分离不同年龄的大鼠胚胎肝干细胞,用含10%优等胎牛血清的H-DMEM培养液培养。利用免疫细胞化学方法在激光共聚焦扫描显微镜下观察该细胞表面抗原的表达情况,进行鉴定。结果两种方法分离的大鼠胚胎肝干细胞体外培养24 h贴壁,5 d左右可长成单层,光镜下为致密圆形细胞,边缘清楚。8 d后细胞铺展,呈上皮样。表达甲胎蛋白(AFP)抗原、细胞角蛋白(CK)18、19。结论所分离的细胞经鉴定为大鼠胚胎肝干细胞,可在体外培养条件下迅速扩增,两种方法所分离培养的肝细胞形态基本一致。
Objective To compare two methods of isolating culture rat embryonic hepatic stem cellls（EHSCs） in vitro and identify the cultured cells. Methods By collagenase perfusion method and cutting digestion to isolate different age rat EHSCs, which are then cultivated by H-DMEM containing 10% fetal bovine serum.To observe the cell surface antigen of rat EHSCs with the confocal laser scanning microscope, using immunocytochemistry for identification. Results With two methods isolated rat EHSCs grow to monolayer after 5 days by cultivation in vitro. They present pykno-round cells and distinct borderline under the light microscope. After 8 days the cells grow like epithelium. EHSCs express AFP antigen,CK 18 and CK19. Conclusion.lusion The cultivated cells are proved to be rat EHSC and can proliferate quickly in vitro. The morphous of rat EHSC isolated by two methods has no differences.
出处
《中国实验诊断学》
2006年第10期1170-1172,共3页
Chinese Journal of Laboratory Diagnosis
基金
国家高技术研究发展计划(863计划)(2004AA205020)
