国产丝裂霉素处理的胎儿成纤维细胞支持具有生殖系转化能力的小鼠胚胎干细胞的分离

Mytomycin Made in China Can Be Used to Isolate Embryonic Stem Cells with Ability in Contributing to Germ Lines

摘要: 应用于胚胎干细胞（embryonic stem cells, ES细胞）培养的国产药物可以降低ES细胞的实验成本。研究中以浙江海正药业生产的丝裂霉素（mitomycin，国药准字H33020786）处理小鼠胎儿成纤维细胞，用于胚胎干细胞建系。并制备饲养层，将小鼠囊胚种植在该饲养层上。4—6天后，挑选形态良好的内细胞团(inner cell mass)来源的克隆在胰酶中进行消化，将消化下来的细胞团块传至新鲜的饲养层上。之后，每2—3天传代一次。结果表明，经该丝裂霉素处理的胎儿成纤维细胞支持具有生殖系嵌合能力的胚胎干细胞的分离。

Abstract: Application of drugs made in China on culture of embryonic stem cells could decrease the cost of ES cell research. In our experiment, mytomycin made in China（Zhejiang Hisun Pharmaceutical Co.,Ltd, authorized H33020786）was used to process feeder cells, from which mouse embryonic stem cells have been isolated and cultured in vitro. When the blastocysts were cultured directly on treated feeder cell layer, the inner cell masses were growing. After 4-6 days culture, ICM-derived outgrowths were separated into clumps, and placed on mitomycin C-treated mouse embryonic fibroblasts feeder layer. Every 2-3 days, passage were made ever since. These ES cells have been proven to have a potential for germ-line contribution.