昆明小鼠原始生殖细胞无血清培养基的筛选  

Selection of serum-free medium for culturing primordial germ cells of Kunming mice

作　　者：董书伟[1] 冯秀亮[2] 段艳萍[2] 杨春荣[1] 杨学义[1] 窦忠英[1] 

机构地区：[1]西北农林科技大学国家干细胞工程技术研究中心陕西分中心,陕西杨凌712100 [2]第四军医大学西京医院实验外科,陕西西安710032

出　　处：《西北农林科技大学学报（自然科学版）》2008年第4期27-31,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30671067);陕西省重大项目(2006K205-G1)

摘　　要：【目的】筛选体外培养昆明小鼠原始生殖细胞（PGCs）的最佳无血清培养基,为昆明小鼠胚胎生殖细胞（EGCs）建系奠定基础。【方法】以妊娠8.5-12.5 d昆明小鼠PGCs为材料,采用无血清培养基,在培养基中添加不同组分,组成A、B、C、D、E、F、G 7种培养基,对昆明小鼠PGCs进行体外培养,分离培养胚胎原始生殖细胞,以筛选出其最佳培养基。【结果】D、E、F组均能得到较多的小鼠PGCs阳性集落。MEF-CM和GR-CM组的效果与细胞因子组相比差异不显著,但降低了培养基的成本。【结论】在无血清培养基中添加0.1 mmol/L RA或使用条件培养基MEF-CM和GR-CM,均有利于小鼠PGCs增殖。经过EGCs鉴定,本试验得到的细胞是PGCs,并且在各组培养基中均能够形成EGCs集落。[Objective] The study was to choose the best serum-free medium for culturing Kunming mice PGCs in vitro. [Method] Groups of mediums, A, B, C, D, E, F and G with different components were used to separate and culture PGCs from 8.5--12.5 days＇ fetuses. [Result] D,E and F groups could culture more positive PGCs colonies. Comparison between groups of MEF-CM and GR-CM with group of cell factors didn＇t show dignificant differences, but they decreased the cost of mediums. [Conclusion] The mediums with 0.1 mmol/L RA or MEF-CM and GR-CM were helpful for proliferation of mouse PGCs. After EGCs were characterized, the cells were proved to be PGCs,and EGCs colonies were obtained in each group of medium.

关 键 词：原始生殖细胞 胚胎生殖细胞 无血清培养基 昆明小鼠 

分 类 号：Q813.11[生物学—生物工程]