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. DOI: 10.7671/j.issn.1001-411X.2013.02.019
    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. DOI: 10.7671/j.issn.1001-411X.2013.02.019
    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. DOI: 10.7671/j.issn.1001-411X.2013.02.019
    Citation: 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. DOI: 10.7671/j.issn.1001-411X.2013.02.019

    猪精原干细胞体外培养条件的优化

    Optimizing the Culture Conditions of Porcine Spermatogonial Stem Cells (PSSCs) in vitro

    • 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.

       

      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.

       

    /

    返回文章
    返回