Preservation and transplantation of porcine testis tissue

Abstract

Grafting of immature mammalian testis tissue to mouse hosts can preserve the male germline. To make this approach applicable to a clinical or field situation, it is imperative that the testis tissue and/or spermatozoa harvested from grafted tissue are preserved successfully. The aim of the present study was to evaluate protocols for the preservation of testis tissue in a porcine model. Testis tissue was stored at 4°C for short-term preservation or cryopreserved by slow-freezing, automated slow-freezing or vitrification for long-term storage. Preserved tissue was transplanted ectopically to mouse hosts and recovered xenografts were analysed histologically. In addition, spermatozoa were harvested from xenografts and cryopreserved. Total cell viability and germ cell viability remained high after tissue preservation. Complete spermatogenesis occurred in xenografts preserved by cooling up to 48 h, whereas spermatogenesis progressed to round spermatids in the xenografts that were frozen–thawed before grafting. Approximately 50% of spermatozoa harvested from xenografts remained viable after freezing and thawing. The in vivo developmental potential of cryopreserved tissue was reduced despite high post-thaw viability. Therefore, it is important to evaluate germ cell differentiation in vivo in addition to cell viability in vitro when optimising freezing protocols for testis tissue.

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers

“…Antibodies against PGP9.5 have been used for immunohistochemical detection of SSCs in several species such as: mouse [15], bovine [16], ram [14], humane [17] porcine [18]. This protein is readily observed in both the cytoplasm and nucleus of gonocytes and spermatogonia in bovine testes.…”

“…In contrast, its signal is relatively faint in cells along the inside of the basement membrane of the mouse seminiferous tubule epithelium. Sertoli cells and spermatogonia also are exhibited weak positive staining for PGP9.5 [18].…”

“…Antibody against this protein has been used for immunohistochemical detection of mouse [15], bovine [16], ram [14], human [17] and porcine [18] spermatogonia. Respecting to the lack of information on goat type A spermatogonial markers and the inadequate information regarding to the isolation, identification, and culture of goat SSCs, and considering the application of transfected SSCs as an efficient tool in production of transgenic animals, the present study was aimed to investigate the identification of prepubertal goat SSCs using polyclonal antibodies against PGP9.5 and c-kit molecular markers.…”

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

“…This technique has been so far successful with a wide range of donor species including hamsters, rabbits, dogs, cats, sheep, goats, horses, bovines, alpacas and monkeys [103] . Work has also shown that testicular tissue pieces can be cryopreserved and later used with this technique in various species [104,105] . Tissue xenografting has represented a step forward in SSC biology that circumvents many of the current limitations the transplantation of germ cell suspensions into the seminiferous tubules of foreign recipient animals.…”

New Approaches of Ovarian Tissue Cryopreservation from Domestic Animal Species

“…Cryopreservation of testicular tissue has been studied since about ten years in animals (Jezek et al, 2001;Jahnukainen et al, 2007;Milazzo et al, 2008;Zeng et al, 2009;Abrishami et al, 2010;Milazzo et al, 2010;Curaba et al, 2011) and human (Bahadur and Ralph, 1999;Bahadur et al, 2000;Guerin, 2005;Revel and Mejia, 2010). It is the only available solution for pre-pubertal boys who must receive a gonadotoxic treatment (eg.…”