Effects of various cryopreservation media and freezing-thawing on the morphology of rat testicular biopsies

Abstract

Currently, testicular sperm extraction is successfully combined with intracytoplasmic sperm injection into the oocyte (ICSI). Several pieces of a testicular biopsy can be frozen and thawed until the ICSI attempt. In this study, the effects of freezing-thawing on the morphology of rat testicular biopsies stored in different cryopreservation media were analysed. Each cryopreservation medium contained glycerol and/or dimethyl sulfoxide (DMSO) as cryoprotectants. In general, both glycerol and DMSO, when applied at moderate concentrations (6-25%), preserved the structure of the seminiferous epithelium. The freezing-thawing procedure had no significant effect on tubular diameter; however, it caused a 'folding' of the lamina propria and notable damage to Sertoli cells, spermatogonia and spermatocytes. Round and elongated spermatids and spermatozoa displayed occasional nuclear damage, vacuolization, and shrinkage/swelling of the cytoplasm. However, the vast majority of these cells maintained their normal structure in nearly all the applied cryomedia. It is concluded that freezing-thawing of testicular biopsies, and the cryopreservation medium, have a significant impact on the structure of the seminiferous epithelium, particularly on its basal compartment.

Effects of different cryoprotectants on the cryopreservation of cattle testicular tissue

“…With sucrose as CPA, testicular seminiferous tubule basal parts were seriously injured. DMSO had been found to be a more suitable CPA than ethylene glycol (EG) for immature mouse and rat testis tissue (Goossens et al, 2008;Jezek et al, 2001). Milazzo et al (2008) reported that freezing testes with DMSO maintained not only immature testicular tissue architecture but also the viability of testicular cells.…”

“…Keros et al (2005) compared the DMSO and sucrose regarding their testicular tissue cryopreservation effect and showed that 10 % DMSO had the best protection effect and 6 % sucrose had the worst protection effect. DMSO has been found to be a more suitable CPA than ethylene glycol (EG) for immature mouse and rat testis tissue (Goossens et al, 2008;Jezek et al, 2001). Yildiz et al (2013) indicated that neonatal mouse testes in DMSO had a higher rate of tissue survival than in PrOH and glycerol after graft.…”

Cryopreservation of Testicular Tissue

“…Later, other detailed studies comparing multiple protocols showed high cell viability with programmed slow-freezing of immature mouse testis tissue using 1.5M DMSO as a cryoprotectant (Milazzo et al, 2008;Traverse et al, 2011). DMSO has also been found to be a more suitable cryoprotective agent than ethylene glycol for immature mouse and rat testis tissue (Goossens et al, 2008;Jezek, 2001). Shinohara et al (2002) reported the birth of mouse offspring from sperm retrieved from cryopreserved pre-pubertal testis tissue with DMSO after transplantation under tunica albuginea of the recipient testes (Shinohara et al, 2002).…”

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.…”