Survival of Bull Sperm Frozen at Different Rates in Media Varying in Osmolarity

The effects of freezing procedures, osmolarity, trehalose, and sucrose on survival of bull sperm in whole milk (WM) and egg yolk–Tris (EYT), semen extenders used worldwide, were studied. Sperm were added to extenders at 25°C, cooled slowly to 5°C, glycerolated, packaged in 0.5-ml straws, and frozen....

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Bibliographic Details
Published in:Cryobiology 1998-11, Vol.37 (3), p.219-230
Main Authors: Liu, Zishu, Foote, Robert H., Brockett, Charles C.
Format: Article
Language:English
Subjects:
Animal cells
Animal productions
Animals
Biological and medical sciences
Biological material (laboratory animals, preparation of biological tracers, etc.)
BIOLOGICAL PRESERVATION
bull sperm
BULLS
Cattle
Cell cultures. Hybridization. Fusion
Cell Survival
CONGELACION
CONGELATION
CONSERVACION BIOLOGICA
CONSERVATION BIOLOGIQUE
CRIOPROTECTORES
CRYOPRESERVATION
Cryopreservation - methods
CRYOPROTECTANTS
CRYOPROTECTEUR
egg yolk–Tris
ESPERMATOZOO
FREEZING
freezing rate
Fundamental and applied biological sciences. Psychology
General aspects
Glucose
Glycerol
In Vitro Techniques
Male
Milk
Molecular and cellular biology
Osmolar Concentration
osmolarity
OSMOTIC PRESSURE
PRESION OSMOTICA
PRESSION OSMOTIQUE
SACCHAROSE
Semen Preservation - methods
Sperm Motility
SPERMATOZOA
Spermatozoa - cytology
Spermatozoa - physiology
SPERMATOZOIDE
SUCROSA
SUCROSE
TAUREAU
Terrestrial animal productions
Time Factors
TORO
TREHALOSA
TREHALOSE
Tromethamine
Vertebrates
whole milk
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Summary:The effects of freezing procedures, osmolarity, trehalose, and sucrose on survival of bull sperm in whole milk (WM) and egg yolk–Tris (EYT), semen extenders used worldwide, were studied. Sperm were added to extenders at 25°C, cooled slowly to 5°C, glycerolated, packaged in 0.5-ml straws, and frozen. Different freezing rates were accomplished in two steps. Straws were transferred from +5°C to nitrogen vapor at temperatures ranging from −10 to −100°C in the first step and to liquid nitrogen in the second step. Straws were thawed in water at 35°C. A substantial decrease in sperm motility occurred between −10 and −20°C, as abrupt nucleation occurred following supercooling to −13°C. To study the interactions between osmolarity × cooling rate, WM and EYT extenders were prepared to yield media measuring 220 to 420 mOsm/L. The optimal first-step range of cooling in the two-step procedure was −30 to −70°C, and the highest proportions of motile sperm after freezing and thawing were 61 to 62 in 260 to 300 mOsm/L WM and 63 to 64% in 300 to 340 mOsm/L EYT, equivalent to the results with the control procedure used commercially. As the cooling rate increased (first step to −100°C) sperm motility was much higher in hypertonic than in hypotonic extenders (P< 0.05), indicating the importance of partial dehydration before rapid cooling. Replacing part of EYT and WM with equivalent solutions (same mOsm/L) of sucrose or trehalose had no appreciable effect. These results provide a basis for further investigating simple freezing systems that might be more effective in preserving bull sperm than those currently available.
ISSN:0011-2240
1090-2392
DOI:10.1006/cryo.1998.2117