title: Evaluation of cryopreservation techniques for bovine embryos.
abstract: A total of 228 embryos was nonsurgically collected from superovulated cows and dehydrated in dimethyl sulfoxide (DMSO) or glycerol by a three-step procedure or a (T.I.T.) timed interval titration procedure. Embryos were loaded in straws, frozen by cooling to -6.0 degrees C at 1.0 degrees C/min, and seeded, followed by cooling to -30 degrees C at 0.3 degrees C/min and to -38 degrees C at 0.1 degrees C/min. At this time the straws were plunged into liquid nitrogen at -195 degrees C. Embryos were thawed in a 27 degrees C or 37 degrees C water bath and rehydrated by a six-step, three-step (sucrose) or one-step (sucrose) procedure. This yielded a 2 x 2 x 2 x 3 factorial treatment structure. Survival was based on development after 12 h in in vitro culture. The only significant single factor affecting survival was the initial quality grade of the embryo. Grades 1 and 2 embryos survived more often than Grade 3 embryos (P < 0.05). Using DMSO as the cryoprotectant resulted in better scores for the post dehydration to post thawing interval (P = 0.02). For both intervals, post dehydration to post thawing and post thawing to post rehydration, the previous quality grade was significant in determining the subsequent quality grade (P < 0.01). At each step of the freeze-thaw process, the embryos became progressively less morphologically intact.
authors.affiliation: Department of Surgery and Medicine Kansas State University Manhattan, KS 66506 USA.
authors.forename: R S
authors.initials: RS
authors.lastname: Prather
authors.forename: M F
authors.initials: MF
authors.lastname: Spire
authors.forename: R R
authors.initials: RR
authors.lastname: Schalles