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大鼠精原细胞的分离纯化
http://www.100md.com 《新疆医科大学学报》 2005年第1期
分离纯化,,大鼠;,精原细胞;,分离纯化;,ckit,1材料和方法,2结果,3讨论,参考文献:
     摘要: 目的: 探讨大鼠精原细胞的分离纯化。 方法: 采用酶消化法制备10 d Wister大鼠睾丸单细胞悬液;Percoll液不连续密度梯度分离精原细胞;用贴壁速度的差异纯化精原细胞。光电镜观察精原细胞的形态结构特征;应用酪氨酸蛋白激酶(ckit)免疫化学鉴定精原细胞。 结果: 精原细胞主要分布在28%~36%(Ⅱ带)间的Percoll梯度;分离纯化的精原细胞形态结构与组织切片上精原细胞一致,其纯度达55.68%;ckit在精原细胞呈阳性表达,在睾丸体细胞呈弱阳性或阴性表达。结论: 利用酶消化法、Percoll液不连续密度梯度及细胞贴壁速度的差异,分离纯化的精原细胞满足体外实验要求;精原细胞表达特异的ckit受体。

    关键词: 大鼠; 精原细胞; 分离纯化; ckit

    Isolation and purification of spermatogonium in rat

    WANG Lin, GU Dantian, GAN Ziming

    (Department of histology and embryology, Xinjiang Medical University,Urumqi 830054,China)

    Abstract: Objective: To study isolation and purification of rat spermatogonium. Methods: Digestions by enzymes were used to prepare germ cell suspension of Wister rat at 10 days;Percoll discontinue density gradient centrifugation and differentiation of velocity of cells sticking to the wall in vitro were used to isolate and purify spermatogonia respectively; Morphological and structural features of spermatogonia in rat at 10 d of age were observed by light and transmission electron microscope; Spermatogonium was identified by ckit (tyrosine protein kinase) immunochemistry. Results: Spermatogonia were mainly distributed in percoll gradient between 28%~36% ......

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