探讨保持胚胎干细胞全能性的体外培养条件  

The Investigation of Cell Culture Conditions Maintained Embryonic Stem Cells as Totipotent Cells

作　　者：邱观婷[1] 黄冰[2] 唐仕波 胡洁 刘志雄 

机构地区：[1]中山医科大学中山眼科中心玻璃体视网膜实验室,广州510060 [2]中山医科大学实验动物中心,广东广州510089

出　　处：《中山医科大学学报》2000年第2期100-103,共4页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：国家自然科学基金!( 3 9770 790 )

摘　　要：【目的】探讨保持胚胎干细胞 (EScells)全能性的体外培养条件。【方法】将ES D3细胞株培养在SNL饲养层细胞和原代小鼠胚胎成纤维细胞饲养层上以及在无饲养层的条件下 ,培养基中加入小鼠白血病抑制因子 (mLIF)或用BuffaloRatLiver(BRL)条件培养基进行培养 ,并对培养细胞进行碱性磷酸酶检测和染色体分析。【结果】ES D3在上述培养体系中能形成正常形态的克隆 ,并且经多次传代后碱性磷酸酶检测为阳性 ,而且核型保持稳定。【结论】说明上述各种培养体系均能保持ES D3的高度未分化状态及正常的二倍体核型。Objective To investigate the culture conditions of embryonic stem(ES) cells which can retain their totipotency. Method ES D3 cells were cultured in the presence of feeder layer cells including SNL cell line and primary mouse embryonic fibroblast (PMEF) and in the absence of feeder layer provided the medium supplemented with purified murine leukemia inhibitory factor(mLIF) or buffalo rat liver (BRL) condition media respectively. The alkaline phosphatase (AKP) test and karyotype analysis were carried out. Results ES D3 could develop normal clone on the above mentioned cell culture systems. Moreover, ES D3 cells within the colonies continued to express AKP over many passages and keep their normal karyotype. Conclusions It indicated that above mentioned culture systems could effectively maintain ES D3 cells undifferentiated state and normal diploid karyotype.

关 键 词：胚胎干细胞 培养基 体外培养条件 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]