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

Implementação da técnica de criopreservação de ovócitos imaturos com posterior descongelação, maturação e fecundação in vitro
Authors: Teixeira, Sofia Margarida Pontes;

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

Abstract

The 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.

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Portugal
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Keywords

Cryopreservation, In Vitro Fertilization, Embryo, Fertilização Animal in vitro, Embriologia Animal, Criopreservação, Vitrificação, Cattle, Vitrification, Bovinos

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selected citations
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This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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