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Plasmocin对人胚胎干细胞支原体污染的杀灭效果

, PP. 783-785

Keywords: 胚胎干细胞,支原体,Plasmocin,bFGF,增殖

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Abstract:

目的人胚胎干细胞(hESCs,humanembryonicstemcells)应用广泛,但来源有限。前期检测发现本室培养的H9hESCs有支原体污染。本研究拟探讨高效支原体杀灭剂plasmocin去除hESCs支原体污染的可能性,及其对hES增殖、分化的影响。方法常规培养hESCs细胞,用切割法进行传代。分别用低(12.5μg/ml)、中(25μg/ml)、高(37.5μg/ml)浓度plasmocin处理hESCs2~3周,用PCR法检测支原体,用结晶紫染色观察hESCs克隆大小,显微镜下观察hESCs分化状况。结果低、中、高浓度plasmocin处理2周均不能杀灭hESCs的支原体,延长高浓度plasmocin处理时间至3周,亦未能杀灭支原体。实验发现,plasmocin呈剂量依赖性抑制hESCs增殖,且高浓度plasmocin可诱导hESCs分化,而高浓度bFGF(40ng/ml)则能部分阻断plasmocin抑制增殖和诱导分化的作用。结论plasmocin未能去除hESCs的支原体污染,可能与支原体产生抗药性及hESCs的克隆生长特性有关。

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