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小鼠精原干细胞分离培养条件的优化

DOI: 10.7671/j.issn.1001-411X.2012.01.018, PP. 87-91

Keywords: 昆明小鼠,精原干细胞,体外培养,MTT法

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Abstract:

采用两步酶消化法获得5~7日龄昆明小鼠睾丸组织的单细胞悬液,比较差速贴壁法和Percoll不连续密度梯度法对精原干细胞(Spermatogonialstemcells,SSCs)的纯化效果,并利用MTT法检测不同生长因子及添加浓度对SSCs体外增殖的影响。结果表明5日龄小鼠和7日龄小鼠的睾丸细胞总数存在较大差异(P<0.05),但SSCs数目无显著差异(P>0.05);与Percoll不连续密度梯度法相比,差速贴壁法获得了较高的SSCs数量和纯化率,其纯化率平均85.06%;MTT法检测结果表明LIF因子的添加对SSCs的增殖具有促进作用,20ng/mLGDNF+20ng/mLbFGF+103U/mLLIF的组合添加是昆明小鼠SSCs的体外培养增殖的最佳组合。

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