In vitro colonization of human spermatogonia stem cells: Effect of patient's clinical characteristics and testicular histologic findings

Objective: To evaluate the effect of the demographic/clinical characteristics of patients and testicular histologic findings on the in vitro colonization of human spermatogonial stem cells (SSCs). In vitro isolation and proliferation of human SSCs has emerged as a suitable method for the enrichment of spermatogonia germ cells. Methods: SSCs were isolated from the testicular biopsies of 47 infertile men with nonobstructive azoospermia and co-cultured with a Sertoli cell monolayer. Age, infertility duration, medical/surgical history, testicular size, and testicular histologic findings were recorded. The patients were divided into 2 groups according to the growth/no growth of human SSC colonies in culture. As the main outcome measure, the number and diameter of germ cell-derived colonies were compared between 2 groups in days 8, 13, and 18 after cultivation with respect to the recorded parameters. Results: No difference was found between the 2 groups regarding the demographic/clinical parameters. Maturation arrest at the premeiotic spermatogonia stage was present in a considerably greater proportion in the group with growth of human SSC colonies compared with the group without growth of human SSC colonies (14 [45.1%] of 31 versus 3 [18.7%] of 16; P <.001) on days 8, 13, and 18 after culture. Maturation arrest at premeiotic SSCs was associated with a greater number and larger diameter of germ cell colonies compared with the maturation arrest at primary spermatocyte and secondary spermatocyte/spermatid stages (P <.001). Conclusion: Infertile men with testicular histologic findings of maturation arrest at the premeiotic spermatogonia stage were seemingly the most appropriate candidates for testicular biopsy and in vitro propagation of human SSCs, regardless of their demographic/clinical characteristics.