The purification of individual male germ cell populations is integral for the molecular and biochemical characterisation of specific spermatogenic phases. Although a number of more contemporary techniques have been developed, velocity sedimentation using the STAPUT method remains as a gold standard for this purpose. The gentle nature of the technique, wherein germ cell subpopulations are separated by sedimentation at unit gravity, results in the isolation of viable and high-purity cells. We provide an updated and simplified step-by-step version of the STAPUT protocol for the purification of mouse male germ cells. As per the original method, the protocol described herein allows for the purification of mouse spermatocyte and round spermatids, however it also allows for successful purification of elongating, and elongated spermatid populations, and is optimised for the preservation of cellular ultrastructure. This method yields sufficient numbers of high-purity cells from one adult mouse for RNA or protein extraction or for immunolocalisation studies.

中文翻译：

---

一种优化的STAPUT方法，用于纯化小鼠精母细胞和精子细胞群体。

单个雄性生殖细胞群体的纯化对于特定生精期的分子和生化表征是必不可少的。尽管已经开发了许多更现代的技术，但使用STAPUT方法进行速度沉降仍然是为此目的的黄金标准。该技术的温和性质（其中生殖细胞亚群通过单位重力下的沉降分离）导致了活细胞和高纯度细胞的分离。我们提供了STAPUT协议的更新和简化的分步版本，用于纯化小鼠雄性生殖细胞。按照原始方法，本文所述的方案可用于纯化小鼠精细胞和圆形精子细胞，但也可用于成功纯化延伸的和延伸的精子细胞群体，并且针对保留细胞超微结构进行了优化。此方法可从一只成年小鼠中产生足够数量的高纯度细胞用于RNA或蛋白质提取或用于免疫定位研究。