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Implementation of immature oocytes by cryopreservation technique and subsquent in vitro maturation and fertilization post-thawing

2011-07-15Master thesis Open access
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dc.contributor.advisorSilva, Joaquim Fernando Moreira da
dc.contributor.advisorFaheem, Marwa Said
dc.contributor.authorTeixeira, Sofia Margarida Pontes
dc.date.accessioned2011-10-17T15:18:08Z
dc.date.available2011-10-17T15:18:08Z
dc.date.issued2011-07-15
dc.descriptionDissertação de Mestrado em Engenharia Zootécnica.por
dc.description.abstractThe aim of this study was to cryopreserve by vitrification by propylene glycol (PROH) and dimethylsulfoxide (DMSO) immature bovine oocytes in straws and to investigate the effects of vitrification on post-thaw oocyte maturation and fertilization. A total of 983 cumulus oocyte complexes were obtained by follicle aspiration from 263 ovaries of cows slaughtered at a local slaughterhouse. Following selection, oocytes with compacted cumulus cells and evenly granulated ooplasm were vitrified using one of the two different solutions with a non vitrified group served as control. The first step vitrification solution contained 10% PROH while the second step solution contained 20% PROH+1M sucrose in a basic media used in group of PROH. Oocytes were matured in N-2-hidroxyethyl piperazine-N-2-ethanosulfonic acid (HEPES) buffered tissue culture medium (TCM) 199 supplemented with 10% FCS, 0.02 IU/ml FSH (Sigma), 1μg/ml E2 (Sigma), 0.15mg/ml Glutamine, 22μg/ml Na-Pyruvate, 10μg/ml Gentamycin, 10μg/ml Streptomycin and 10μg/ml Nistamycin. for 24 h at 38 ◦C in a humidified atmosphere of 5% CO2 in air. Oocytes were then fertilized and cultured. The numbers of developed embryos observed were 98 (70.1%), 73 (59.9%), and 80 (67.7%) in groups Control, PROH and DMSO, respectively, without co-culture. With co-culture were observed 141 (69.5%), 54 (61.7%), and 45 (49%), also in Control, PROH and DMSO. Developed embryos rate from control, without co-culture, was not statistically different when compared with PROH and DMSO (p< 0.05), but with co-culture present statistical differences with DMSO. However, better results were obtained in DMSP group compared to PROH, without co-culture. The lowest number of developed embryos was obtained in DMSO group, with co-culture. Immature bovine oocytes can be vitrified in straws, but success differs with the cryoprotectant and without co-culture.por
dc.identifier.urihttp://hdl.handle.net/10400.3/1241
dc.language.isoengpor
dc.subjectCattlepor
dc.subjectCryopreservationpor
dc.subjectEmbryopor
dc.subjectIn Vitro Fertilizationpor
dc.subjectVitrificationpor
dc.subjectBovinospor
dc.subjectCriopreservaçãopor
dc.subjectEmbriologia Animalpor
dc.subjectFertilização Animal in vitropor
dc.subjectVitrificaçãopor
dc.titleImplementation of immature oocytes by cryopreservation technique and subsquent in vitro maturation and fertilization post-thawingpor
dc.title.alternativeImplementação da técnica de criopreservação de ovócitos imaturos com posterior descongelação, maturação e fecundação in vitropor
dc.typemaster thesis
dspace.entity.typePublication
rcaap.rightsopenAccesspor
rcaap.typemasterThesispor

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DCA - Dissertações de Mestrado / Master Thesis