| 碱性磷酸酶细胞化学法检测体外培养精原细胞 |
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| 引用本文： | 李莲军,陆峻波,施江滨,苗永旺,廖红芬,李德雪.碱性磷酸酶细胞化学法检测体外培养精原细胞[J].云南农业大学学报,2004,19(3):318-321. |
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| 作者姓名： | 李莲军 陆峻波 施江滨 苗永旺 廖红芬 李德雪 |
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| 作者单位： | [1]云南农业大学动物科学技术学院，云南昆明650201 [2]解放军军需大学动物科学技术系，吉林长春130062 |
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| 摘 要： | 分别培养2日龄、6日龄及12日龄小鼠生精上皮细胞48～96 h,并进行碱性磷酸酶（AP）细胞化学染色,以及波形蛋白（vimentin）和c-Kit受体蛋白免疫细胞化学染色,观察细胞形态及染色特性,以研究不同日龄小鼠生精上皮细胞在体外培养时形态及AP特性,建立一种简便可行的检测精原细胞的方法。结果,管周细胞为不规则形状,呈vimentin+反应、c-Kit－反应和AP+反应;幼稚型及正成熟的支持细胞（Sertoli cell）为大小不等的不规则形状,呈vimentin+反应、c-Kit－反应和AP－反应;6 日龄小鼠精原细胞为圆球形或近似圆球形,呈vimentin－反应和c-Kit+反应。生殖细胞中,前精原细胞（性原细胞）和精原细胞均呈AP+反应,部分精母细胞也呈AP+反应。研究结果表明,在没有前精原细胞及精母细胞的培养体系中,可以借助细胞形态和AP活性两项指标检测其中的精原细胞。
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| 关 键 词： | 精原细胞 碱性磷酸酶 体外培养 小鼠 |
| 文章编号： | 1004-390X(2004)03-0318-04 |
| 收稿时间： | 2003-6-18 |
Detection of Cultured Spermatogonia with Alkaline Phosphatase Activity in vitro |
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| LI Lian-jun,LU Jun-bo,SHI Jiang-bin,MIAO Yong-wang,LIAO Hong-fen,LI De-xue.Detection of Cultured Spermatogonia with Alkaline Phosphatase Activity in vitro[J].Journal of Yunnan Agricultural University,2004,19(3):318-321. |
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| Authors: | LI Lian-jun LU Jun-bo SHI Jiang-bin MIAO Yong-wang LIAO Hong-fen LI De-xue |
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| Institution: | LI Lian-jun~1,LU Jun-bo~1,SHI Jiang-bin~1,MIAO Yong-wang~1,LIAO Hong-fen~1,LI De-xue~2 |
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| Abstract: | The dissociated seminiferous epithelial cells from 2 to 12 days of postpartum mice were cultured for 48～96 h, and the morphological characterization and the expression of vimentin, c-Kit, AP activity of them were examined cytochemically and immunocytochemically, respectively to establish a simple and reliable method of detecting spermatogonia in a culture system. The peritubular cells of all ages examined have a spread irregular outline, and demonstrated vimentin positive reaction, c-Kit negative reaction and AP positive reaction. Immature and matuting Sertoli cells of all ages examined took the shape of irregularity and demonstrated vimentin positive reaction, c-Kit negative reaction and AP negative reaction. Spermatogonia from 6 dpp mice shown vimentin negative reaction and c-Kit positive reaction, and were spheral outline. Gonocytes and spermatogonia all were AP positive reaction. Some of spermatocytes were AP positive reaction. It is concluded that spermatogonia in a culture system without gonocytes and spermatocytes in existence can be detected by using AP activity combined with cell morphological criteria. |
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| Keywords: | spermatogonium alkaline phosphatase culture in vitro mouse |
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