| 摘要: |
| 为探索提高精原干细胞分离和体外培养效率的技术方法，以建立长期稳定的动物精原干细胞培养体系，对动物精原干细胞的分离纯化和体外培养的研究进展进行综述。以“Spermatogonial stem cell，Isolation，Culture，Enrichment，in vitro”和“精原干细胞、分离、培养和体外扩增”为检索词，查阅精原干细胞相关文献，并进行归纳整理。精原干细胞分离主要有机械分离法和酶消化法，纯化精原干细胞的方法有差速贴壁法、密度梯度离心法、免疫磁珠法、流式细胞法等，目前大动物精原干细胞的富集常使用酶消化法、差速贴壁法和Percoll梯度离心法相结合。精原干细胞体外培养需模拟体内生境，通常在基础培养基中添加血清、饲养层细胞和各类生长因子等，培养条件会影响精原干细胞培养的效果。小鼠精原干细胞已建立较完善的培养体系，而人和多数大动物还缺乏完善的精原干细胞体外培养体系。 |
| 关键词: 精原干细胞 A型精原细胞 分离 纯化 体外培养 |
| DOI：10.11841/j.issn.1007-4333.2017.04.10 |
| 投稿时间：2016-03-09 |
| 基金项目:西北农林科技大学博士科研启动费（Z111021303），引进人才科研启动费（Z111021203） |
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| Progress on the research of spermatogonial stem cells cultured in vitro |
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SUN Xiuzhu, ZHU Yanjiao, DU Lijuan, WANG Liqiang, KANG Mengyang, WANG Shuhui
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| (College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China) |
| Abstract: |
| In order to explore the technology and methods to improve the efficiency of isolation and culturing of spermatogonial stem cells,and to establish long-term stable animal spermatogonial stem cell culture system,the isolation,purification and culture in vitro of animal spermatogonial stem cells.Literatures were searched using keywords of "spermatogonial stem cell,isolation,culture,enrichment,in vitro" both in Chinese and in English,and the relevant literatures were collated.The results show that:Spermatogonial stem cells can be isolated by mechanical separation and enzymatic digestion;Purification of spermatogonial stem cells are differential plating,density gradient centrifugation,magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS);At present,large animals spermatogonial stem cells are commonly enriched by enzyme digestion,differential adherence method and Percoll gradient centrifugation;Culture in vitro of Spermatogonial stem cells should be simulated in vivo,which usually added serum,feeder cells and various growth factors in the basal medium;Conditions affect the results of spermatogonial stem cell culture.It was found that a perfect system has been established for mouse spermatogonial stem cells culture in vitro,while there is still a lack of fine spermatogonial stem cells culture system for human and the majority of the large animals. |
| Key words: spermatogonial stem cells type A spermatogonium isolation purification in vitro culture |
