Pachytene spermatocytes can achieve meiotic process in vitro

Highly enriched pachytene spermatocytes prepared from adult rats by centrifugal elutriation were tested for their capacity to enter the meiotic process when cocultured with 20-day old rat Sertoli cells. This was traced by phase-contrast microscopy and by DNA flow cytometry. We also performed a Northern blot analysis using the mouse protamine I cDNA as a probe, the expression of which being restricted to spermatids. Our results demonstrate that pachytene spermatocytes cocultured with Sertoli cells developed into spermatids. The number of pachytene spermatocytes entering meiosis was affected neither by growth factors nor by hormones. However these later were required in long term cocultures for the maintenance of cell integrity and viability.