Cryopreservation of spermatozoa from cynomolgus monkeys (Macaca fascicularis)

Abstract

Three egg-yolk diluents, which have been used successfully in cryopreservation of human spermatozoa, were compared for their ability to protect macaque semen against cryodamage. TEST (Tes + Tris + egg yolk), TEST with 20% skim milk (TSM), and egg yolk-citrate (EYC), each with 3 or 5% glycerol were compared using 12 ejaculates from 6 male cynomolgus macaques. Computer-aided analysis of sperm motion was used to determine the percentage motility (%M), curvilinear velocity (VCL), and linearity (LIN) of spermatozoa after thawing. The supravital stain Hoechst 33258 and a fluoresceinated pea lectin were used to determine the % of viable spermatozoa with intact acrosomes. TSM and TEST were superior to EYC in terms of % M and of % viable, acrosome-intact spermatozoa. TSM and TEST produced equivalent VCL and LIN values, while EYC had clearly reduced VCL and LIN. There were no interactions between diluent and glycerol level. The 3% glycerol level gave superior results to 5% glycerol for %M. EYC, which is widely used for cryopreservation of human spermatozoa, was not suitable for cynomolgus monkey semen. Artificial insemination with semen cryopreserved in TSM resulted in a healthy, full-term infant.

In vitro fertilization‐embryo transfer in nonhuman primates: The technique and its applications

“…The first live birth following insemination with frozen-thawed sperm was achieved in the gorilla (Douglass, 1981). Similar results were reported recently in the chimpanzee (Gould and Styperek, 1989) and the cynomolgus monkey (Tollner et al, 1990). Sperm cryopreservation procedures differ in cooling and freezing rates, buffer systems, and extenders, but glycerol serves as the common cryoprotectant.…”

Aging in male primates: reproductive decline, effects of calorie restriction and future research potential

“…Studies of semen characteristics in aging nonhuman primates have been cross-sectional, often with single or short time-point measures and small sample sizes (Platz et al 1980;Tollner et al 1990;Schaffer et al 1992;VandeVoort et al 1993;Morrell 1997;Yeoman et al 1997;Ramesh et al 1998;Ji et al 2001). Some studies have focused on testosterone levels in male rhesus macaques, with somewhat contradictory results, similar to the human data (Mattison et al 2001;Black and Lane 2002;Mattison et al 2003;Roth et al 2004).…”

Delay of Ovulation Due to Diets Containing Levonorgestrel in Cynomolgus Monkeys (<i>Macaca fascicularis</i>)

“…Although reproductive technology for monkeys, such as artificial insemination and embryo transfer has been improved [4,5,[10][11][12], control of the ovarian cycle has not been established. There are a few reports showing that progestins are useful for control of ovulation in monkeys.…”