Epub 2005 Dec 15.Schäfer-Somi S1, Kluger S, Knapp E, Klein D, Aurich C.Author information1Centre for Artificial Insemination and Embryo Transfer, University of Veterinary Medicine, Vienna 1210, Austria. sabine.schaefer@vu-wien.ac.atAbstractThe aims of the present study were, to assess the effects of semen centrifugation, two different diluents and two different freezing methods on post-thaw semen quality in canine semen, and to elucidate the interdependence of these parameters. For this purpose, the sperm-rich fractions of ejaculates from 12 healthy male beagles were divided into four aliquots. Two aliquots were centrifuged and resuspended with two TRIS-egg yolk based extenders: with Uppsala and Gill extender (Gill). The diluents differed in the concentration of glycerol and in the admixture of Equex STM paste (Nova Chemical Sales Inc., Scituate, MA, USA). Diluted semen was frozen either in a styrofoam box or with a computerized freezing machine and an optimized freezing curve (IceCube 1,810;

The change in temperature inside the straws was measured during the freezing procedure. Thawed semen samples were assessed for motility and viability (SYBR-14/PI) using the computer assisted sperm analyzer SpermVision (Minitüb, G) and a modified triple staining technique (flow cytometry). Deep freezing in the machine resulted in better motility and viability than in the box. The combination centrifugation-Uppsala extender-machine was superior to all other combinations, which was most evident after storage at +5 degrees C for 7 h (motility: 53.1%, viability: 64.9%). Post-thaw longevity and progressive motility were significantly improved by the use of the here introduced freezing curve. This was shown to be partly caused by less pronounced fluctuations of temperature inside the straws when compared to box-freezing.PMID: 16359725 DOI: 10.1016/j.theriogenology.2005.10.022 [PubMed - indexed for MEDLINE] Publication TypesComparative StudyMeSH TermsAnimalsCell SurvivalCryopreservation/instrumentationCryopreservation/methodsCryopreservation/veterinary*Cryoprotective Agents/pharmacology*Dogs/physiology*Flow Cytometry/veterinaryMaleSemen Preservation/instrumentationSemen Preservation/methodsSemen Preservation/veterinary*Specimen Handling/methodsSpecimen Handling/veterinarySperm Count/veterinarySperm MotilitySpermatozoa/physiology*Time FactorsSubstancesCryoprotective AgentsFull Text SourcesElsevier ScienceMiscellaneousNCI CPTAC Assay PortalNCI CPTAC Assay PortalNCI CPTC Antibody Characterization ProgramNCI CPTC Antibody Characterization Program

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