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Updated: Nov 6, 2025
Sperm Collection of Differential Quality Using Density Gradient Centrifugation
Published on: November 29, 2018
Quality Improvement of Post-Thawed Stallion Epididymal Spermatozoa with Single Layer Centrifugation
F C Cunha Dos Santos1, J M Morrell2, M M Nunes3
1Programa de Pos Graduacao Medicina Animal: Equinos, Faculdade de Veterinaria, Universidade Federal do Rio Grande do Sul, Rio Grande do Sul, Brazil. carlini@portoweb.com.br.
Background:
Epididymal sperm cryopreservation represents the ultimate option to preserve spermatozoa of valuable stallions.
Objective:
The study aims to evalute whether single layer centrifugation (SLC) prior to cryopreservation or after post-thawing improves the quality of stallion epididymal sperm.
Materials And Methods:
Epididymal sperms of stallions were harvested (N=20). Sperm samples were subjected to treatments: conventional centrifugation, SLC prior to cryopreservation (SLC-PC) or SLC post-thaw (SLC+). All samples were cryopreserved, thawed and evaluated. SLC+ were thawed, single layer cenrifuged and resuspended in freezing extender (SLC+F) or cooling extender (SLC+C). Total motility, progressive motility, morphology, mitochondrial functionality, membrane integrity and DNA integrity were evaluated.
Results:
SLC-PC and SLC+F yielded higher total motility, while SLC+F yielded the highest progressive motility. Mitochondrial functionality was significantly higher in all SLC groups. Membrane integrity was higher in SLC-PC. The percentage of morphologically normal spermatozoa was higher in SLC-PC and SLC+F.
Conclusion:
SLC prior to cryopreservation or post-thaw improves the quality of stallion epididymal spermatozoa. When SLC is performed post-thaw, freezing extender is the best medium to resuspend the pelleted semen.
