Long-term coculture of spermatogonial stem cells on sertoli cells feeder layer in vitro

To construct a cell model of spermatogonial stem cells (SSCs) cocultured on Sertoli cells feeder layer in vitro, and study the proliferation characteristics of SSCs.

Sertoli cells and SSCs were separated from testes of 14-15 days and 6 days KM mice respectively by two-step enzyme digestion. SSCs were seeded on the Sertoli cells layer at 5 days in culture. The clones of SSCs on the sertoli cells layer were detected, and cast-off cells in culture medium were counted.

SSCs began to proliferate and differentiate 24 hours after being cultured on the Sertoli cells layer, and there were a few paired (Ap) cell clones. With more time of culture, the number of Ap cell clones decreased gradually, meanwhile the number of aligned (Aal) cell clones increased, then Aal cell colonies retained stable quantity after 120 hours in culture, it could retain (51.2 +/- 5.8) days under the condition of the culture medium being changed every 4 or 5 days.

SSCs can proliferate colonially on the feeder layer of Sertoli cells, and retain stable morphous and quantity. SSCs cultured on Sertoli cell feeder layer provide a cell model for studying SSCs biological behavior and interruptions of drugs or toxins on spermatogenesis in vitro. Coculture