Preparation and short-term culture of enriched populations of guinea pig spermatocytes and spermatids.

ABSTRACT

In order to study acrosome biogenesis in vitro, the authors developed techniques for the isolation and culture of enriched populations of guinea pig spermatogenic cells using a modification of the technique of Romrell et al (1976). The modifications include changes in the medium, enzyme concentrations, cell loads for gradient separations, and sedimentation times. Three major cell populations were pooled Pachytene spermatocytes (PS 2.0 x 10(7), 80-85% pure), round spermatids, (RS 1.2 x 10(8), 80-85% pure), and condensing spermatids (CS 2.0 x 10(8), 50-60% pure, contaminated with residual bodies). The ultrastructural properties of the isolated cells appeared similar to those of cells in situ. PS were 3-4 times more active than RS and 10 times more active than CS in synthesizing proteins. These experiments demonstrate that highly enriched populations of guinea pig spermatogenic cells can be isolate, and that these cells can be cultured in the presence of radioactive precursors for studies of protein synthesis during spermatogenesis.