Use of density centrifugation for delayed cryopreservation of stallion sperm: perform sperm selection directly after collection or after storage?
Equipment for cryopreservation of stallion sperm is not always available. In such cases, diluted semen can be shipped to a facility for later cryopreservation. The aim of this study was to evaluate if selection of sperm via density centrifugation yields higher survival rates when cryopreservation is to be delayed (i.e. carried out after 1 day of storage at 5°C). Two-layer iodixanol as well as single-layer Androcoll density centrifugation were tested and compared with samples prepared with standard centrifugation. Special emphasis was placed on comparing centrifugation on the day of semen collection with centrifugation after 1-day refrigerated storage. Sperm morphology and motility as well as membrane and chromatin integrity were evaluated before and after centrifugation. Sperm motility and membrane integrity were also assessed after cryopreservation. It was found that both two- and single-layer density centrifugation processing resulted in higher percentages of morphologically normal and motile sperm with higher membrane and chromatin integrity, as compared to standard centrifugation or diluted samples. Differences were only in the order of magnitude of 5%. Recovery rates after density centrifugation were only approximately 30-40%. When cryopreservation was carried out after 1-day refrigerated storage, centrifugation processing of sperm directly after semen collection resulted in higher percentages of plasma membrane intact sperm post-thaw as compared to performing centrifugation processing of stored sperm just prior to cryopreservation. No significant differences in progressively motile sperm post-thaw were seen. Taken together, for delayed cryopreservation, it is best to perform density centrifugation directly after collection rather than immediately prior to cryopreservation.
Clinic for Horses - Unit for Reproductive Medicine, University of Veterinary Medicine Hannover, Hannover, Germany.
A
A
Heutelbeck
H
H
Oldenhof
K
K
Rohn
G
G
Martinsson
J M
JM
Morrell
H
H
Sieme
D000177:Acrosome / Q000502:physiology; D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000662:veterinary*; D002843:Chromatin / Q000502:physiology; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D006736:Horses; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology* / Q000648:ultrastructure; D013997:Time Factors
D002843:Chromatin