Protocol for ex vivo culture of adult mouse testis tissue using a gel-based approach

Testicular function relies on tightly regulated cell-cell interactions, including Sertoli-Leydig cell communication and Sertoli cells nurturing germ cells. Here, we present a 48-h ex vivo culture protocol for adult mouse testis tissue using a gel-based approach. We describe experimental design, dissection, and examination of treatment-induced outcomes, in particular, stimulatory effects on germ cells. We outline procedures for quantifying apoptosis and germ cell proliferation in the tissue and measurements of reproductive hormones and paracrine factors secreted to the culture medium. For complete details on the use and execution of this protocol, please refer to Andreassen et al.¹.