In vitro production of fertile sperm from murine spermatogonial stem cell lines

Abstract

spermatogonial stem cells (ssCs) are the only stem cells in the body that transmit genetic information to the next generation. The long-term propagation of rodent ssCs is now possible in vitro, and their genetic modification is feasible. However, their differentiation into sperm is possible only under in vivo conditions. Here we show a new in vitro system that can induce full spermatogenesis from ssC lines or any isolated ssCs. The method depends on an organ culture system onto which ssCs are transplanted. The settled ssCs form colonies and differentiate up into sperm. The resultant haploid cells are fertile, and give rise to healthy offspring through micro-insemination. In addition, the system can induce spermatogenesis from ssCs that show spermatogenic failure due to a micro-environmental defect in their original testes. Thus, an in vitro system is established that can be used to correct or manipulate the micro-environmental conditions required for proper spermatogenesis from murine ssC lines.

Testis tissue explantation cures spermatogenic failure in c-Kit ligand mutant mice

“…Recently, we succeeded in inducing complete spermatogenesis in mice using an organ culture method (3)(4)(5). In this culture system, germ cells developed from spermatogonial stem cells through mitotic differentiation of spermatogonia, meiosis in spermatocytes, and morphological transformation in haploid spermatids, to sperm.…”

Evaluation of ovarian and testicular tissue cryopreservation in children undergoing gonadotoxic therapies

“…Cryopreservation of testicular tissue followed by subsequent transplantation resulted in restoration of testicular function in a variety of animal hosts, including mice, rabbit and chicken [18,40,43]. A recent murine study has succeeded in maturation of spermatogonia in testicular tissue in vitro resulting in the generation of viable offspring, in addition to the survival of testicular tissue for 2 months in vitro [38]. These data support the possibility of maturing spermatogonia in human pediatric testicular tissue in vitro, which may be an important consideration in prepubertal males.…”

818 Long-term ex vivo maintenance of testis tissues producing fertile sperm in a microfluidic device

“…The culture medium was drawn through the outlet by the syringe pump at 0.05 μ L/min. Tissues of the AG group were cultured on agarose stands (1.5% w/v) placed in wells of a 12-well culture plate (CELLSTAR ® Tissue Culture Plates, Greiner Bio-One) [3][4][5] . Each gel was loaded with 1-3 testis tissue fragments.…”

“…Although there are several ways to culture a tissue fragment or small organ, the golden standard has long been the so-called interphase method in which samples are placed at the interphase between the culture medium and a gas layer 2 . We adopted this classical organ culture method for culturing mouse testis tissues, and succeeded in inducing complete spermatogenesis, from spermatogonial stem cells up to fertility-competent sperm formation [3][4][5] . Nonetheless, the overall efficiency and duration of the spermatogenesis were far from comparable to those observed in vivo.…”