| 湖羊精原干细胞体外培养 |
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| 引用本文： | 陈晓宇,朱志伟,于福先,宋雪梅,潘建治.湖羊精原干细胞体外培养[J].浙江农业学报,2010,22(5):580. |
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| 作者姓名： | 陈晓宇 朱志伟 于福先 宋雪梅 潘建治 |
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| 作者单位： | 浙江省农业科学院 畜牧兽医研究所,浙江 杭州 310021 |
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| 基金项目： | 浙江省自然科学基金，浙江省农业科学院科技创新提升工程 |
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| 摘 要： | 研究了湖羊精原干细胞(SSCs)体外培养方法,湖羊睾丸曲精细管两步酶消化法制备细胞悬液,Percoll分离后比较SSCs纯化方法、FCS以及GDNF对SSCs体外培养与集落碱性磷酸酶(AKP)染色的影响。结果显示,虽然盘化法纯化的SSCs在集落形成时间与集落数上显著低于差速贴壁法(P<0.05),但是盘化法所得集落的AKP阳性率显著高于差速贴壁法(P<0.05)；添加1% FCS的培养基可显著降低集落形成的时间,增加集落的数目与AKP阳性率(P<0.05),但集落形成时间与AKP阳性率在5%与1% FCS之间无显著变化(P>0.05)；20 ng/mL GDNF处理组内集落数与AKP阳性率显著高于10 ng/mL GDNF(P<0.05),但集落形成时间与AKP阳性率与30～40 ng/mL GDNF的处理组无显著差别(P>0.05)。
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| 关 键 词： | 精原干细胞(SSCs) 体外培养 集落 湖羊 |
Culture of Hu-sheep spermatogonial stem cells in vitro |
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| CHEN Xiao-yu,ZHU Zhi-wei,YU Fu-xian,SONG Xue-mei,PAN Jian-zhi.Culture of Hu-sheep spermatogonial stem cells in vitro[J].Acta Agriculturae Zhejiangensis,2010,22(5):580. |
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| Authors: | CHEN Xiao-yu ZHU Zhi-wei YU Fu-xian SONG Xue-mei PAN Jian-zhi |
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| Institution: | Institute of Animal Husbandry and Veterinary, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China |
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| Abstract: | To develop a series of practical culture methods of Hu-sheep spermatogonial stem cells (SSCs) in vitro for future studies on transgenesis, seminiferous tubule cells were isolated through a two step enzyme digestion from dissociated testis, and the cell suspension was classified by percoll gradient. The effects of different purification methods, FCS and GDNF on SSCs culture in vitro and alkaline phosphatase (AKP) staining were investigated. The results were as follows: (1) The time of colonies formation and colonies numbers from panning method were significantly lower than that from differential plating (P<0.05), in contrast, AKP positive rates from panning method significantly increased (P<0.05). (2) The colonies formation time, numbers and AKP positive rates significantly increased when the medium supplemented with 0.5% BSA and 1% FCS (P<0.05), however, there were no significant difference in the colonies formation time and AKP positive rates between 1% and 5% FCS (P>0.05). (3) The colonies number and AKP positive rates were significantly higher under 20 ng/mL GDNF than that under 20 ng/mL GDNF (P<0.05), but there were no significant difference in colonies formation time and AKP positive rates between 20 ng/mL GDNF and 30-40 ng/mL GDNF (P>0.05)． |
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| Keywords: | spermatogonial stem cells (SSCs) culture in vitro colonies Hu-sheep |
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