In vitro production of porcine zygotes using intracytoplasmic injection of vitrified sperm

CLAUDIA ARRAZTOA; CLARA BACA CASTEX; GABRIEL MARTIN ALVAREZ; PABLO DANIEL CETICA; DEBORA NEILD

REPRODUCTION IN DOMESTIC ANIMALS (1990)

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2017 vol. 52 p. 775 - 780

0936-6768

Theobjectivesofthisstudyweretoevaluateifvitrifiedporcinespermatozoaareabletomaintaintheircapacitytoproducezygotesinvitrousingintracytoplasmicsperminjection(ICSI)andtoevaluatethezygotedevelopmentintwoinvitroatmosphericconditions:5%CO2andtri-gas.Agroupofporcineoocytesmaturatedinvitrowereinjectedwithvitrified-warmedsperm(treatmentgroup)andanothergroup,withspermdilutedandconservedat17°C(controlgroup).Toevidenceparthenogeneticactivation,someoocytesweresubmittedtoaShamtest.TheinjectedoocyteswereculturedinG1mediumat38°C,100%humidityand5%CO2ortri-gas.Nosignificantdifferences(p >.05)wereobservedinembryodevelopmentbetweentheoocytesinjectedwithvitrified-warmedsperm(31.8%;36/113),andthoseinjectedwithsemendilutedandconservedat17°C(35.5%;32/90),whenculturedin5%CO2 or under tri-gasatmosphere(42.9%;39/91vs.34.2%;26/76,respectively).Nosignificantdifferences(p>.05)wereobservedinthepercentageofpronuclei(PN)obtainedbetween5%CO2andtri-gas,withineachtreatmenteither.Ofthe52oocytessubmittedtotheShamtest,onlytwopresentedafemalePN(activation)indicatingthatthePNobservedinthetreatmentgroupwereaproductoffertilizationandnotparthenogeneticactivation.Toconclude,porcinespermvitrifiedusingspheres,ataconcentrationof5×106spermatozoa/mlinTALPmediumwith1%bovineserumalbumin(BSA),conservecondensedandintactchromatincapableofproducingearlyembryodevelopmentuptothepronuclearstage.