Isolation of Sertoli, Leydig, and spermatogenic cells from the mouse testis

被引:116
|
作者
Chang, Yao-Fu [1 ]
Lee-Chang, Jennifer S. [1 ]
Panneerdoss, Subbarayalu [1 ]
MacLean, James A., II [2 ]
Rao, Manjeet K. [1 ]
机构
[1] Univ Texas Hlth Sci Ctr San Antonio, Greehey Childrens Canc Res Inst, San Antonio, TX 78229 USA
[2] So Illinois Univ, Sch Med, Dept Physiol, Carbondale, IL 62901 USA
基金
美国国家卫生研究院;
关键词
spermatogenesis; testis; Sertoli cell; Leydig cell; germ cell; spermatocyte; spermatid; elutriation; ROUND SPERMATIDS; GENE; PURIFICATION; EXPRESSION;
D O I
10.2144/000113764
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A thorough understanding of the events during mammalian spermatogenesis requires studying specific molecular signatures of individual testicular cell populations as well as their interaction in co-cultures. However, most purification techniques to isolate specific testicular cell populations are time-consuming, require large numbers of animals, and/or are only able to isolate a few cell types. Here we describe a cost-effective and timesaving approach that uses a single protocol to enrich multiple testicular cell populations (Sertoli, Leydig, and several spermatogenic cell populations) from as few as one mouse. Our protocol combines rigorous enzymatic digestion of seminiferous tubules with counter-current centrifugal elutriation, yielding specific testicular cell populations with >80%-95% purity.
引用
收藏
页码:341 / +
页数:3
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