Generation of tail bearing sperm-like cells from in vitro spermatogenesis of farming goat testis

Abstract

In vitro spermatogenesis offers a therapeutic modality for the treatment of male infertility associated with pre-or post-meiotic barriers to the spermatogenesis process. The present study reports the production of tail-bearing sperm-like cells from in vitro spermatogenesis of farming goat testis in an air-liquid inter-phase culture. The testicular fragments (1-2 mm3) were placed on agarose scaffolds at air-liquid interphase and cultured in vitro for 60 days. Proliferating germ cell colonies were visible after 45-55 days of culture. Differentiation of cells to spermatogonia and spermatocytes was observed from the 40th day onwards. Tail bearing sperm-like elongated cells, with apical acrosome, were detected from the 45th day onwards. The differentiating germ-line cells were confirmed by stage-specific expression of positive markers (VASA, DAZL, SYCP1, SYCP3, C-Kit, POU5F1 and SOX-2). Histological sections of the cultured testicular tissue further confirmed the presence of tail-bearing sperm-like cells in the lumen of seminiferous tubules which was absent in non-cultured tissue. In conclusion, our study suggests that the air-liquid interphase organ culture might be useful in vitro generation of sperm in farming goats. The study could have a significant impact on the livestock management of the farmers and could improve their dependency on orthodox and natural breeding management.