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

Isolation of Sertoli, Leydig, and spermatogenic cells from the mouse testis.
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DOI:
10.2144/000113764
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发表时间:
2011-11
期刊:
影响因子:
2.7
通讯作者:
Rao MK
Rao MK
中科院分区:
工程技术4区
文献类型:
--
作者:
Chang YF;Lee-Chang JS;Panneerdoss S;MacLean JA 2nd;Rao MK

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为了彻底了解哺乳动物精子发生过程中发生的事件,需要研究单个睾丸细胞群体的特定分子特征以及它们在共培养中的相互作用。然而,大多数分离特定睾丸细胞群的纯化技术是耗时的,需要大量的动物,和/或只能分离少数细胞类型。在这里,我们描述了一种经济有效且节省时间的方法,它使用一个单一的方案来丰富只有一只小鼠的多个睾丸细胞群(支持细胞、间质细胞和几个生精细胞群)。我们的方案结合了生精小管的严格酶消化和逆流离心洗脱,产生了纯度为80%-95%的特定睾丸细胞群。
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.
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