鸡精原干细胞分离培养初步研究.pdfVIP

安徽农业科学,Journal of Anhui Agri .Sci .2008 ,36( 34) :14973 - 14975 责任编辑 李玮 责任校对 张士敏 鸡精原干细胞分离培养初步研究 1 1 * 1 2 2 1 1 采克俊 , 刘莉 , 张易祥 , 于立颖, 丁海雷, 丁志丽, 谢小丹 ( 1. 湖州师范学院生命科学学院, 浙江湖州313000 ;2 . 扬州大学动物科学与技术学院, 江苏扬州 225009) 摘要 [ 目的] 建立禽 类精原干细胞(SSCs) 体外培养 系统。[ 方法] 以25 日龄广西土 鸡为试材, 通过 睾丸石蜡切 片和 HE 染 色, 了解 SSCs 发育情况, 用两步酶消化 法分 离获得生精上皮细胞, 比较不 同温度、接种密度对鸡SSCs 体外培 养的影响, 并对SSCs 的集落进行 染 色鉴 定。[ 结果] 结果显表 明,25 日龄 时SSCs 已经形成, 但还未分化 。37 ℃适宜作为鸡SSCs 的培养温度, 5 ×105 个/ ml 适宜作为鸡SSCs 原代 混合培养的接种密度 。经碱性磷酸酶和糖原染 色鉴定, 体外培养的鸡SSCs 仍保持未分化的特性。[ 结论] 该研 究为禽 类SSCs 体外培养 的基础资料提供参考 。 关键词 鸡; 精原干细胞; 分 离; 培养; 鉴定 中图分类号 S831 .1 文献标识码 A 文章编号 0517 - 6611(2008) 34 - 14973 - 03 Preli minary Study on the Isolation and i n vitr o Culture of Chicken Sper matogonial Stem Cells CAI Ke-j un et al ( School of Life Sciences , Huzhou Teachers college , Huzhou , Zhej iang 313000) Abstract [ Obj ective] The ai mof study was to found in vitro culture systemof SSCs for birds kind . [ Method] 25- day- old male chickens were used to iso- late seminiferous epithelial cells by sequential two- step enzyme digestion . The sper matogonial stemcells ( SSCs) development stage was evaluated by histological examination after he matoxylin- eosin staining . The effects of different culture temperatures and concentration on pri mary culture of chicken SSCs were evaluated , and the SSCs colonies were characterized by staining . [ Result] The results showed that by 25- days , SSCs , as the most advanced germcells , were present , but 5 undifferentiated . The temperatures at 37 ℃and seeding at a concentration of 5 ×10 cells/ ml were suitable for chicken SSCs culture . During in vitro culture , the colon