Porcine sperm vitrification I: cryoloops method.
The aims of this study were to evaluate porcine sperm vitrification in cryoloops, with and without two different cryoprotectants and assess two warming procedures. Extended (n = 3; r = 4) and raw (n = 5; r = 2) semen was diluted in media without and with cryoprotectants (4% dimethylformamide and 4% glycerol) to a final concentration of 20 × 106spermatozoa ml-1and vitrified using the cryoloops method. Two warming procedures were evaluated: rapid method (30 s at 37°C) and an ultra-rapid method (7 s at 75°C, followed by 30 s at 37°C). Total motility (phase contrast), sperm viability (6-carboxifluorescein diacetate and propidium iodide stain), membrane function (hypo-osmotic swelling test), acrosome integrity (phase contrast), chromatin condensation (toluidine blue stain) and chromatin susceptibility to acid denaturation (acridine orange stain) were evaluated before and after vitrification and analysed using Friedman's test. In all media, the only seminal parameters that were maintained after vitrification were chromatin condensation and integrity. Vitrification of porcine spermatozoon using cryoloops, both in the presence or absence of cryoprotectants and independent of the warming procedure used, permits conservation of sperm chromatin condensation and integrity. It would be interesting to further verify this by producing porcine embryos using vitrified spermatozoon with intracytoplasmic sperm injection.
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.|Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Arraztoa
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.|Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Miragaya
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.|Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Chaves
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.|Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Trasorras
Cátedra de Estadística, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Gambarotta
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Péndola
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.|Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
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Neild
D000177:Acrosome / Q000648:ultrastructure; D000818:Animals; D001947:Breeding; D002462:Cell Membrane / Q000502:physiology; D002470:Cell Survival; D002843:Chromatin / Q000737:chemistry / Q000502:physiology; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents; D006358:Hot Temperature; D008297:Male; D055101:Semen Analysis / Q000662:veterinary; D012662:Semen Preservation / Q000295:instrumentation* / Q000379:methods* / Q000662:veterinary*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology / Q000648:ultrastructure; D034421:Sus scrofa / Q000235:genetics
D002843:Chromatin; D003451:Cryoprotective Agents
cryoloop; cryopreservation; porcine; sperm; vitrification