[三种培养基中小鼠精原干细胞的增殖情况]

[The proliferation profile of mouse spermatoganial stem cells in three types of culture media].

作者信息

Wang Qing-zhong

机构信息

College of Bio-Engineering, Weifang University, Shandong Province Weifang, 261061, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2009 May;25(2):286-8.

PMID:21189574

Abstract

AIM

To establish an in vitro long-term culture system of mouse Spermatogonial stem cells (SSCs).

METHODS

Three types of serum-free culture media, namely, DMEM/F12, KSR (KnockoutM Serum Replacement) and StemPro-34 SFM, to which the same growth factors including GDNF, soluble GFRalpha1 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

StemPro-34 SFM serum-free medium sustains the proliferation of mouse SSCs in vitro.

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

使用三种无血清培养基，即DMEM/F12、KSR（敲除血清替代物）和StemPro-34 SFM，均添加相同的生长因子，包括胶质细胞源性神经营养因子（GDNF）、可溶性GFRα1和碱性成纤维细胞生长因子（bFGF），并使用小鼠胚胎成纤维细胞（MEF）饲养层培养通过差异贴壁选择从幼鼠睾丸富集的小鼠SSCs。通过形态学检查干细胞活性，并通过逆转录-聚合酶链反应（RT-PCR）和免疫细胞化学分析检测SSCs的标记基因表达。