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.PDFVIP

动 物 学 研 究 2007, Dec. 28(6) ：654－658 CN 53-1040/Q ISSN 0254-5853 Zoological Research 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 * 魏著英，邵 华 ，刘东军，旭日干 （内蒙古大学哺乳动物生殖生物学与生物技术教育部重点实验室，内蒙古 呼和浩特 010021 ） 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）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。 关键词：国产丝裂霉素；小鼠胚胎成纤维细胞；小鼠胚胎干细胞 中图分类号：Q593+ .4 文献标识码：A 文章编号：0254-5853 （2007 ）06-0654-05 Mytomycin Made in China Can Be Used to Isolate Embryonic Stem Cells with Ability in Contributing to Germ Lines * WEI Zhu-ying, SHAO Hua , LIU Dong-jun, BOU Shor-gan （The key laboratory for mammalian reproductive biology and biotechnology, ministry of education. Hohhot 010021, China） 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 ） 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 fo