Spermatogenesis is a series of cellular processes that leads to the development of motile, elongate sperm cells. Mitotic expansion of spermatogenic stem cells is followed by two meiotic cell divisions that yield haploid round spermatids which then transform from a spherical form into an elongate, highly polarized form. In Drosophila, spermatogenesis takes place within encapsulating cysts that contain spermatogenic cells. Spermatogenic cysts were isolated and grown in culture over the course of 96 hours. Cultures were treated with buthionine sulfoximine (BSO), glutathione (GSH), insulin, and GSH+insulin in order to test the effects of these agents on cyst viability. The addition of glutathione and exogenous insulin to cultured spermatogenic cysts each appeared to have a positive effect on early spermatogenic cyst survival in vitro at some timepoints. The addition of GSH+insulin together had no significant effect on early spermatogenic cyst survival in vitro. Oxidative stress induced by BSO resulted in a significant decrease and/or complete loss of specific early spermatogenic cyst types and the abnormal development of elongating cysts in culture. This culture system offers the opportunity for high-resolution analysis of spermatogenic processes not previously possible.