Greehey Children's Cancer Research Institute, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Biotechniques. 2011 Nov;51(5):341-2, 344. doi: 10.2144/000113764.
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.
要深入了解哺乳动物精子发生过程中的事件，需要研究单个睾丸细胞群体的特定分子特征及其在共培养物中的相互作用。然而，大多数分离特定睾丸细胞群体的纯化技术既耗时又费力，需要大量的动物，而且/或者只能分离少数几种细胞类型。在这里，我们描述了一种经济高效且省时的方法，该方法使用单个方案从仅一只小鼠中富集多种睾丸细胞群体（支持细胞、间质细胞和几种精原细胞群体）。我们的方案将生精小管的严格酶消化与逆流离心洗脱相结合，得到纯度>80%-95%的特定睾丸细胞群体。
