Postmortem recovery and cryopreservation of spermatozoa from the vas deferens of rhesus macaques (Macaca mulatta)

Abstract

To determine whether sperm derived form the vas deferens could be retrieved and successfully cryopreserved, testes were collected from 20 rhesus macaques (Macaca mulatta). The males ranged in age from 3 to 19 years with an average of 8.5 years. No sperm was obtained from three animals which were younger than 4 years. The remaining 17 samples contained sperm with an average sperm cell number of 421.8 ±88.7 × 106 and an average motility of 72.8 ± 4.4 %. After 24 h of culture in TALP medium at 37° C in 5% CO2 and 95% air, the overall motility decreased significantly in all samples regardless of treatment. Freezing in TEST-yolk buffer containing 6% (vol/vol) glycerol had a significant effect on sperm reducing the immediate post-thaw motility to 42.4 % in non-treated samples. Treatment with db-cAMP and caffeine further reduced sperm motility after 4 hr in fresh sperm (72.8 vs 50.4%), but increased in sperm that had been frozen (14.0 vs 23.2%). The age of the male did not influence sperm concentration or grade, but proved to be a significant factor in determining motility of frozen-thawed treated sperm with lower motility found in samples from older males. Overall the study demonstrates that motile sperm can be obtained from post-mortem males, although subsequent studies will have to determine whether the quality is sufficient to facilitate its use in assisted reproduction.

Age-Associated Pathology in Rhesus Macaques (Macaca mulatta)

“…Cryopreservation studies have shown that older rhesus macaques (~19 years) have fresh sperm concentrations and grade (quality) comparable to that of young males but significantly reduced motility in thawed cryopreserved samples. 35 …”

“…Cryopreservation studies have shown that older rhesus macaques (*19 years) have fresh sperm concentrations and grade (quality) comparable to that of young males but significantly reduced motility in thawed cryopreserved samples. 35 The aged rhesus macaque prostate has been extensively evaluated due to the need for models of benign prostatic hyperplasia and prostatic carcinoma. Studies and case reports of rhesus macaque prostatic pathology have identified stromal hyperplasia, cystic hyperplasia, squamous metaplasia, focal glandular pleomorphism, nonsuppurative prostatitis, rare single adenocarcinomas, and more commonly, benign basal cell hyperplasia.…”

Artificial Insemination in Veterinary Science

“…Epididymal spermatozoa have been harvested from a variety of species like cats (Filliers et al, 2008;Hermansson & AxnÈr, 2007), dogs (Garcia-Macias et al, 2006;Hewitt et al, 2001;Nothling et al, 2007;Ponglowhapan et al, 2006), rats (Yamashiro et al, 2007), horses (Braun et al, 1994;Bruemmer, 2006;Heise et al, 2011;Johnson & Coutinho da Silva, 2008;Melo et al, 2008), cattle (Goovaerts et al, 2006;Martins, Rumpf, Pereira, & Dode, 2007), pigs (Ikeda et al, 2002), sheep (Garcia-Macias et al, 2006), goats (Blash, Melican, & Gavin, 2000), red deer (Fernandez-Santos et al, 2006;Garcia-Macias et al, 2006;MartÌnez-Pastor et al, 2006), Spanish Ibex , African buffalo (Herold et al, 2006), North American buffalo (Lessard at al., 2009) and monkeys (Goff et al, 2009;Ng et al, 2002). Pregnancies and offspring after AI with epididymal spermatozoa have been produced amongst others in horses (Barker & Gandier, 1957;Heise et al, 2010;Papa et al, 2008), dogs (Hori, Hagiuda, Kawakami, & Tsutsui, 2005) and Spanish Ibex (SantiagoMoreno et al, 2006).…”

Optimization of Ethylene Glycol Concentrations, Freezing Rates and Holding Times in Liquid Nitrogen Vapor for Cryopreservation of Rhesus Macaque (Macaca mulatta) Sperm

“…The optimal freezing rate for sperm cryosurvival should be low enough to avoid IIF but fast enough to minimize the solution effects. In the conventional freezing method, rhesus sperm are usually frozen in LN 2 vapor in a Styrofoam box after being loaded into cryostraws [2,5,10,16,20,21]. Therefore, the variations in penetrating CPA concentrations in sperm freezing medium and the freezing rates resulting from varied distances between cryostraws and the LN 2 surface affect the cryosurvival of rhesus sperm.…”