| 小鼠精原干细胞分离培养条件的优化 |
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| 引用本文： | 张秀娟,李莉,卫恒习,白银山,师如意,张守全.小鼠精原干细胞分离培养条件的优化[J].华南农业大学学报,2012,33(1):87-91. |
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| 作者姓名： | 张秀娟 李莉 卫恒习 白银山 师如意 张守全 |
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| 作者单位： | 华南农业大学动物科学学院,广东广州,510642 |
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| 基金项目： | 国家973计划，国家支撑计划 |
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| 摘 要： | 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(Spermatogonial stem cells,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,20 ng/mL GDNF+20 ng/mLbFGF+103 U/mL LIF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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| 关 键 词： | 昆明小鼠 精原干细胞 体外培养 MTT法 |
| 收稿时间： | 4/6/2011 12:00:00 AM |
The Optimization of Isolation and Culturing of Spermatogonial Stem Cells in Mouse |
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| ZHANG Xiu-juan,LI Li,WEI Heng-xi,BAI Yin-shan,SHI Ru-yi and ZHANG Shou-quan.The Optimization of Isolation and Culturing of Spermatogonial Stem Cells in Mouse[J].Journal of South China Agricultural University,2012,33(1):87-91. |
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| Authors: | ZHANG Xiu-juan LI Li WEI Heng-xi BAI Yin-shan SHI Ru-yi and ZHANG Shou-quan |
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| Institution: | (College of Animal Science,South China Agricultural University,Guangzhou 510642,China) |
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| Abstract: | Testicular cells of 5-7 days postpartum Kunming mouse were collected by two-step enzymatic digestion method.Effects of two different purification methods known as different adherence selection and percoll discontinuous density gradients were compared,whereas the effects of different growth factors on spermatogonial stem cells(SSCs) proliferation were investigated as well by MTT assay.The results showed that there was significant difference in the numbers of total testes cells acquired between 5 days postpartum pups and 7 days postpartum pups(P<0.05),yet the numbers of SSCs had no significant difference between the two groups(P>0.05).The purification method of different adherence selections attained higher SSCs numbers and purification ratio than Percoll method,with the ratio of 85.06%.By means of MTT assay,the results showed that LIF factor could promote SSCs proliferation,and the 20 ng/mL GDNF+20 ng/mL bFGF+103 U/mL LIF was the best supplementation combination in Kunming mouse SSCs in vitro. |
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| Keywords: | Kunming mouse spermatogonial stem cells in vitro culture MTT assay |
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