title: The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spermatozoa: a comparison of two controlled-rate cooling machines.
abstract: Spermatozoa from three species, bovine, ovine, and porcine, were frozen using standard techniques in two controlled-rate cooling machines, a commercial instrument and a custom-built device. Ice crystallisation was induced mechanically by touching the straws with a pre-cooled rod. The sperm samples were stored 24h, and then thawed rapidly and evaluated for motility, viability, and acrosomal integrity in the membrane-intact population. The custom-built controlled-rate cooling machine proved significantly better at all cooling rates for all species. This was particularly evident for the ram and the boar spermatozoa. In general, -30 or -50 degrees C/min were better than -1 degrees C/min, with a slight advantage being evident for -30 degrees C/min. However, this became very apparent for boar spermatozoa. It is clear that the higher cooling rates are necessary for successful freezing of spermatozoa from these species, and that careful control of the cooling rate is essential for maximal recovery of viable and functional cells. This is best achieved when the cooling profile is controlled from within a dummy sample.
authors.affiliation: Department of Veterinary Basic Sciences, Royal Veterinary College, Royal College Street, London NW1 0TU, UK.
authors.forename: S
authors.initials: S
authors.lastname: Kumar
authors.forename: J D
authors.initials: JD
authors.lastname: Millar
authors.forename: P F
authors.initials: PF
authors.lastname: Watson
mesh_terms: D000818:Animals; D002417:Cattle / Q000502:physiology*; D002470:Cell Survival; D015925:Cryopreservation / Q000295:instrumentation*; D008297:Male; D012662:Semen Preservation / Q000379:methods*; D012756:Sheep / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine / Q000502:physiology*