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【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 in vitro culture conditions for maintaining the pluripotency of embryonic stem cells (ESCs). 【Method】 The ES D3 cell line was cultured on feeder layer of primary and secondary mouse embryonic fibroblast feeder cells and feeder-free conditions, and the mouse leukemia inhibitory factor (mLIF) was added to the culture medium or supplemented with Buffalo Rat Liver (BRL) conditioned medium, and alkaline phosphatase detection and chromosome analysis on the cultured cells. 【Result】 ES D3 could form normal clones in the above culture system and tested positive for alkaline phosphatase after multiple passages, and the karyotype remained stable. 【Conclusion】 These results indicate that all the above culture systems can maintain the highly undifferentiated ES D3 and normal diploid karyotypes.