| 猪精原干细胞体外培养条件的优化 |
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| 引用本文： | 师如意,卫恒习,李莉,白银山,张秀娟,吴珍芳,张守全.猪精原干细胞体外培养条件的优化[J].华南农业大学学报,2013,34(2):224-229. |
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| 作者姓名： | 师如意 卫恒习 李莉 白银山 张秀娟 吴珍芳 张守全 |
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| 作者单位： | 1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室, 2 山西医科大学 基础医学院, 省部共建教育部细胞生理学重点实验室,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室, 3 广东省昆虫研究所,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室,;1 华南农业大学 动物科学学院, 广东省农业动物基因组学与分子育种重点实验室, |
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| 基金项目： | 国家973计划（2011CB944202,2010CB945001,2009CB941601）；国家科技支撑计划（2011BAD19B03） |
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| 摘 要： | 以出生后1～5 d的长白公猪为主要研究对象,采用两步酶消化法获取睾丸细胞悬液,用Percoll不连续密度梯度法对猪精原干细胞(PSSCs)进行纯化,以碱性磷酸酶(AP)染色鉴定;利用MTT法检测2种细胞因子——胶质细胞源性神经营养因子(GDNF)和白血病抑制因子(LIF)对PSSCs发育的影响.结果表明:用Percoll不连续密度梯度法获得的PSSCs占睾丸总细胞数的(8.19±0.83)%,AP染色的阳性率为(87.47±2.90)%,表明该方法可有效获取PSSCs;MTT法检测表明,20 ng.mL-1 GDNF+1 000 U.mL-1 LIF的组合添加对PSSCs体外增殖的效果最好.
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| 关 键 词： | 猪 精原干细胞 体外培养 |
| 收稿时间： | 2012/2/21 0:00:00 |
Optimizing the Culture Conditions of Porcine Spermatogonial Stem Cells(PSSCs) in vitro |
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| SHI Ruyi,WEI Henxi,LI Li,BAI Yinshan,ZHANG Xiujuan,WU Zhenfang,ZHANG Shouquan.Optimizing the Culture Conditions of Porcine Spermatogonial Stem Cells(PSSCs) in vitro[J].Journal of South China Agricultural University,2013,34(2):224-229. |
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| Authors: | SHI Ruyi WEI Henxi LI Li BAI Yinshan ZHANG Xiujuan WU Zhenfang ZHANG Shouquan |
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| Institution: | 1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, 2 Key Laboratory of Cellular Physiology, Ministry of Education, Preclinical Medical College, Shanxi Medical University,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, 3 Guangdong Entomological Institute,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University,;1 Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, |
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| Abstract: | The testicular cells of Landrace piglets aged from 1 to 5 days were isolated by two-step enzymatic digestion method. The Percoll discontinuous density gradients method was used to purify the porcine spermatogonial stem cells (PSSCs). The effects of glial cell-line derived neurotrophic factor (GDNF) and leukemia inhibitory factor (LIF) on PSSCs proliferation were investigated by MTT method. The results showed that the method for PSSCs isolation and purification were effective. By means of MTT assay, the results indicated that LIF and GDNF factor could promote PSSCs proliferation, and the combination of 20 ng·mL-1 GDNF and 1 000 U·mL-1 LIF showed the strongest effects. |
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| Keywords: | porcine spermatogonial stem cells (PSSCs) in vitro culture |
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