Optimized protocol for isolation of germ cells from mouse testis by centrifugal elutriation

Abstract

Germline development is challenging to study due to the diversity of cell types in mammalian testis. Here, we present an optimized protocol, namely centrifugal elutriation, that allows the simultaneous isolation of mouse germ cells at different stages with high purity within ∼2 h. This approach exploits the JE-5.0 centrifugal elutriation system that fractionates cells based on differential sedimentation gravity. We herein provide the optimized parameters and procedures for isolation of elongating spermatids, round spermatids, and pachytene spermatocytes from adult mouse testes. For complete details on the use and execution of this protocol, please refer to Bao et al. (2018).

Keywords: Cell isolation; Developmental biology; Genetics.