摘要
目的:建立小鼠精原干细胞(SSCs)的体外长期培养体系。方法:用分别添加了等量的胶质细胞源神经营养因子(GDNF)、可溶性GFRα1和bFGF的DMEM/F12、KSR和StemPro-34 SFM三种无血清培养基和MEF饲养层分别培养经差异贴壁分选富集的小鼠SSCs,通过形态观察、标志基因的RT-PCR和免疫细胞化学分析检测其SSCs本原。结果:DMEM/F12与KSR可支持小鼠SSCs在体外存活6-7 d,而StemPro-34 SFM能能维持SSCs体外增值一个月。结论:StemPro-34 SFM支持小鼠SSCs的体外增殖。
To establish an in vitro long-term culture system of mouse Spermatogonial stem ceils （SSCs）. Methods： Three types of serum-free culture media, namely, DMEM/F12, KSR （Knockout^TM Serum Replacement） and StemPro-34 SFM, to which the same growth factors including GDNF, soluble GFRα1 and bFGF were added equally, and MEF（mouse embryonic fibroblast） feeder layer were used to culture mouse SSCs enriched from pup mice testes through differential adherence selection. The activity of stem cells was examined morphologically, and the marker gene expression of SSCs was detected by RT-PCR and immunocytochemical analysis. Results： The activity of SSCs cultured in DMEM/F12 and KSR serum-free media was only maintained for 6-7 days. However, the StemPro-34 SFM medium could maintain the proliferation of cultured SSCs nearly one month. Conclusion： StemPro34 SFM serum-free medium sustains the proliferation of mouse SSCs in vitro.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2009年第2期286-288,共3页
Chinese Journal of Applied Physiology
基金
山东省自然科学基金资助(Y2007079)
潍坊市科技发展计划项目(2007028)
关键词
精原干细胞
无血清培养基
培养
小鼠
spermatogonial stem cells
serum-free culture medium
culture
mouse
