268 GERM CELL DEVELOPMENT IN EQUINE TESTIS TISSUE XENOGRAFTED INTO MICE

Abstract

Grafting of testis tissue from immature animals under the back skin of immunodeficient mice results in complete spermatogenesis, albeit with different levels of efficiency in different species. While spermatogenesis develops comparably to that in the donor species in xenografts from pigs, sheep and goats, spermatogenic differentiation is less efficient in testis tissue from cats and bulls. Testicular maturation was significantly accelerated in rhesus monkey testis grafts whereas timing was similar to that in the donor species in cats and bulls. The objective of this study was to investigate if grafting of immature horse testis tissue would result in spermatogenesis in a mouse host. Small fragments of testis tissue (about 1 mm³) from four sexually immature colts (2-week-old Standardbred, 5- and 8-month-old ponies, 10-month-old Warmblood) were grafted under the back skin of castrated male immunodeficient mice (n = 5, 5, 10 and 5 recipient mice, respectively). Histological examination of the testis xenografts was performed between 14 and 50 week post-transplantation. Weight of the seminal vesicles in the host mouse was recorded as an indicator of bioactive testosterone produced by the xenografts. At the time of grafting, the seminiferous cords of the donor testis tissue form 2-week-, 5-month- and 8-month-old colts contained only immature Sertoli cells and gonocytes. No spermatogenic differentiation occurred in xenografts from the 2-week-old colt and testosterone production was minimal. Pachytene spermatocytes were observed in testis grafts from the 5- and 8-month-old donors from 14 weeks onward. Spermatogenesis did not proceed through meiosis in grafts from the 5-month-old donor. Recipient mice carrying xenografts from the 8-month-old donor received exogenous gonadotropins (equine chorionic gonadotropin and human chorionic gonadotropin, 10 I.U./day for 2 months, beginning 14 weeks after grafting) and condensing spermatids were observed by 35 weeks after grafting. In donor tissue from the 10-month-old colt, pachytene spermatocytes were present in about 50% of tubules at the time of grafting. After 14 weeks, xenografts showed fewer differentiated germ cells than the donor tissue. However, at 35 weeks after grafting, condensing spermatids were observed, indicating that differentiated germ cells were initially lost and spermatogenesis was subsequently reinitiated. In all castrated host mice where spermatogenic differentiation occurred, the weight of the seminal vesicles was restored to pre-castration values showing that xenografts were releasing bioactive testosterone. These results indicate that horse spermatogenesis can occur in a mouse host albeit with low efficiency. Testicular maturation was not accelerated. In most cases, spermatogenesis appeared to become arrested at meiosis. The underlying mechanisms of this spermatogenic arrest require further investigation. Although equine testis xenografts produced testosterone, supplementation of exogenous gonadotropins might support post-meiotic differentiation.

This work was supported by USDA 03-35203-13486.