Long-term cultures of testicular biopsies from boys with cryptorchidism: Effect of FSH and LH on the number of germ cells

Abstract

Background: A long-term culture system of testicular biopsies from boys with undescended testes was established to evaluate the effect of gonadotrophins on germ cell survival and growth. Methods: Biopsies from 25 boys with cryptorchidism, aged 1.1-9.9 years, were obtained at the time of surgery. Each biopsy was subdivided into fragments and cultured for 1 or 3 weeks in a testis culture medium without gonadotrophins or supplemented with FSH (50 IU/l) or LH (5 IU/l), or a combination of FSH (50 IU/l) plus LH (5 IU/l). The survival of the germ cells was evaluated by calculating the ratio of spermatogonia and gonocytes per cross-sectioned testicular tubule, the S/T ratio. Results: All cultured fragments maintained their overall morphology. A significantly reduced S/T ratio was observed in the fragments cultured in the presence of LH (P = 0.006), independently of the culture period. The mean tubular diameter in fragments cultured for 1 week with hormones was significantly larger than that of the controls (P < 0.0002). All cultured fragments had a larger tubular diameter than the uncultured fragments (P = 0.0002). Conclusions: This culture system supports survival of spermatogonia for at least 3 weeks. LH either alone or in combination with FSH results in a significantly reduced S/T ratio.