大鼠生精细胞分离方法的研究

Isolation of Spermatogenic Cells from Rat Testis

目的:建立一种简单、有效的哺乳动物睾丸生精细胞分离方法。方法:采用研磨-Percoll法、单一酶-研磨-Percoll法、组合酶法制备成熟期前大鼠睾丸生精细胞,分析比较其细胞总数、分离所需时间和细胞纯度,以及检测细胞培养前后的存活率。结果:组合酶法得到的睾丸生精细胞总数最多、分离所需时间最短;获得的细胞含杂质最少、细胞存活率最高;细胞培养48h后的存活率高,与其他两种分离方法比较有显著性差异(P<0.05)。结论:用组合酶法能在较短时间内获得数量较多的、比较纯化和存活率较高的生精细胞。

Objective: To establish a simple and effective method to isolate spermatogenic cells from rat testis. Methods: Three kinds of cell isolation methods, namely, grind - percoll method, single enzyme - grind - percoll method, and combination of enzymatic digestion, were used to obtain spermatogenic cells from rat testicular tissues. The total number of cell, the isolating time and the cell livability were observed. Results: The combination of enzymatic digestion obtained the largest number of cell and the highest percentage of living cells and consumed the shortest time among the three methods (P < 0.05). The survival rate of the cells isolated by combination of enzymatic digestion was the highest after cultured in vitro for 48h (P < 0.05). Conclusions : The combination of enzymatic digestion is an effective method to obtain spermatogenic cells from rat testicular tissue.