The aim of the present work was to quantitate the temporal and stage-specific effects of follicle-stimulating hormone (FSH) and testosterone on the proliferation and differentiation capacities of the human seminiferous epithelium. Seminiferous tubule fragments were kept in culture for 28 days and 5-bromo-2'-deoxyuridine incorporation was used to determine cell proliferation. Data demonstrated a gradual loss of germ cells during the culture period, no decrease in Sertoli cell numbers, and maintenance of the general architecture of the seminiferous tubules. Both FSH and testosterone increased germ cell survival, spermatogonia proliferation, and germ cell differentiation, especially during the first week of culture. At the end of the first week, differentiation of spermatocytes was observed, especially when 50 IU/L FSH and 1 µmol/L testosterone were used. In conclusion, using this methodology, it was possible to quantify germ cell proliferation and differentiation, in a reproducible way, with results compatible with the timing of human spermatogenesis in vivo.