ISOLATION AND BIOCHEMICAL-STUDIES OF ENRICHED POPULATIONS OF SPERMATOGONIA AND EARLY PRIMARY SPERMATOCYTES FROM RAT TESTES
ISOLATION AND BIOCHEMICAL-STUDIES OF ENRICHED POPULATIONS OF SPERMATOGONIA AND EARLY PRIMARY SPERMATOCYTES FROM RAT TESTES
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DOI：
10.1095/biolreprod34.1.195
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发表时间：
1986-02-01
影响因子：
3.6
通讯作者：
MEISTRICH, ML
中科院分区：
文献类型：
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作者：
BUCCI, LR;BROCK, WA;MEISTRICH, ML
描述了一种从单一年龄大鼠获得几种高度富集的睾丸细胞类型群体的方法。在酶促去除间质细胞后，制备来自未成熟大鼠睾丸的单细胞悬浮液。通过离心淘洗，根据大小将细胞分离为4个级分（散装制剂）或8个级分（分析制剂）。这些淘析器级分通过Percoll梯度中的平衡密度离心进一步分离。以这种方式，2 × 10 - 15的群体107个A型精原细胞（51%纯度）3 ×107个B型精原细胞（纯度76%），5 × 10 - 6个。107个偶线期/早粗线期精母细胞（56%纯度），3 ×107个中粗线期精母细胞（70%纯度），和4 ×50只未成年大鼠处死后6 h内可获得107个支持细胞（纯度89%）。通过细胞学涂片检查确定的纯度通过库尔特体积和流式细胞术DNA测定进行验证。这些分离方法用于获得细胞群，用于表征精子发生早期阶段高迁移率族蛋白的水平和合成。
A method to obtain several highly enriched populations of testis cell types from rats of a single age is described. Single cell suspensions from immature rat testes were prepared after enzymatic removal of interstitial cells. Cells were separated on the basis of size into four fractions (bulk preparations) or eight fractions (analytical preparations) by centrifugal elutriation. These elutriator fractions where further separated by equilibrium density centrifugation in Percoll gradients. In this manner, populations of 2 .times. 107 type A spermatogonia (51% purity) 3 .times. 107 type B spermatogonia (76% purity), 5 .times. 107 zygotene/early pachytene spermatocytes (56% purity), 3 .times. 107 midpachytene spermatocytes (70% purity), and 4 .times. 107 Sertoli cells (89% purity) could be obtained from 50 immature rats within 6 h after killing. Purities, determined by examination of cytologic smears, were verified by Coulter volume and flow cytometric DNA determinations. These separation methods were used to obtain cell populations for characterization of levels and synthesis of high mobility group proteins in the early stages of spermatogenesis.