碱性磷酸酶细胞化学法检测体外培养精原细胞  

Detection of Cultured Spermatogonia with Alkaline Phosphatase Activity in vitro

作　　者：李莲军[1] 陆峻波[1] 施江滨[1] 苗永旺[1] 廖红芬[1] 李德雪[2] 

机构地区：[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

出　　处：《云南农业大学学报》2004年第3期318-321,共4页Journal of Yunnan Agricultural University

摘　　要：分别培养2日龄、6日龄及12日龄小鼠生精上皮细胞48～96h,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(Sertolicell)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。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.

关 键 词：精原细胞 碱性磷酸酶 体外培养 小鼠 

分 类 号：S865.1[农业科学—野生动物驯养]