Benchmarks defining high-quality sperm in the common marmoset (Callithrix jacchus).

Abstract

Background: Common marmosets (Callithrix jacchus) are increasingly recognized as valuable nonhuman primates (NHPs) for biomedical research due to their small size and short reproductive cycle and lifespan relative to other NHP species. Maximizing the utility of captive research marmosets, including genetically manipulated animals, will require the use of assisted reproductive techniques (ART) including manipulation, storage, and sharing of marmoset sperm. Here, we identify characteristics of high-quality semen samples and validate a simple method for selecting high-quality sperm.

Methods: Computer-assisted sperm analysis (CASA) was used to evaluate sperm quality in semen samples collected from 44 marmosets and assessed the use of the swim-up method for the selection of high-quality sperm was also tested in half the samples as a potential means to optimize in vitro fertilization or intrauterine insemination.

Results: For each reference parameter, samples at or below the 5th percentile were categorized as abnormal sperm, while those above the 5th percentile were considered to be normal. Among normal samples, those at or above the 50th percentile were categorized as high-quality. High-quality semen samples exhibited the following characteristics: semen volume ≥ 30 µL; sperm count ≥ 10⁷/ejaculate; total motility ≥ 35%; and normal morphology ≥ 5%. Sperm isolated by swim-up exhibited superior sperm progressive motility (19.7% ± 4.5 vs. 5.6% ± 2.1; P = 0.01) and normal morphology (13.1 ± 1.59 vs. 7.65 ± 1.1; P < 0.001) compared with unselected sperm.

Conclusion: This study defines robust, statistically supported reference values for evaluating marmoset semen samples to assist with the identification of optimal sperm donors and the selection of high-quality sperm samples for assisted reproduction. Ultimately, these reference values combined with a validated selection method will contribute to consistent standards for the international sharing of genetically diverse and/or gene-edited marmoset sperm for research and reproduction.