Improved vitrification method allowing direct transfer of goat embryos

Guignot, Florence; Bouttier, Alain; Baril, Gérard; Salvetti, Pascal; Pignon, P.; Beckers, Jean‐François; Touzé, Jean-Luc; Cognié, Juliette; Traldi, A. S.; Cognie, Y.; Mermillod, Pascal

doi:10.1016/j.theriogenology.2006.02.040

Cited by 45 publications

(52 citation statements)

References 41 publications

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“…These data are in agreement with Guignot et al (2006), who found kidding rates of 56% and 27% when compared vitrified goat embryos with or without sucrose, respectively. These same authors also observed a tendency to increase the embryo survival rate using sucrose.…”

Section: Resultssupporting

confidence: 93%

“…After last bath, the embryos were loaded into 0.25 ml straws with 20-30 µl of vitrification solution and separated by two bubbles of the same solution (approximately 90 µL, each). Then the straws were sealed, identified and immediately immersed in liquid nitrogen (GUIGNOT et al, 2006).…”

Section: Vitrification and Warmingmentioning

confidence: 99%

“…However, the need for high concentrations of cryoprotectants can cause damage to cells by osmotic stress and chemical toxicity (PAPADOPOULOS et al, 2002) and to reduce the ability to reach the glassy state. Seeking to decrease these obstacles in vitrification process, several protocols have been developed using different cryoprotectants, such as ethylene glycol (GUIGNOT et al, 2006), DMSO (HONG et al, 2007), glycerol and propylene glycol (WERLICH et al, 2006). Furthermore, the addition of macromolecules such as sucrose or trehalose has been reported with the function of promoting a significant osmotic effect and it is also used for rehydration of embryos after warming (HURTT, 1999;ISACHENKO et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

et al. 2015

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This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P < 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality. Key words: Cryopreservation, embryo, ewe, multiple ovulation, sugar ResumoEste estudo objetivou avaliar a eficácia da adição de sacarose na solução de vitrificação de embriões ovinos produzidos in vivo. Foram selecionadas 40 ovelhas da raça Dorper as quais foram superovuladas. Imediatamente antes da colheita de embriões por laparotomia, uma laparoscopia foi realizada para verificar a resposta superovulatória. O lavado recuperado foi submetido à procura e avaliação de embriões e estes foram divididos em dois grupos experimentais, onde os embriões do Grupo Controle foram submetidos ao protocolo tradicional de vitrificação e os embriões do Grupo Sacarose a um protocolo modificado de vitrificação com sacarose. Após a descongelação, os embriões foram novamente divididos considerando a remoção (Indireto) ou não (Direto) do crioprotetor. A qualidade embrionária foi classificada como embriões de graus I (excelente ou bom), II (regular), III (pobre) e IV (morto ou degenerado). Foi também verificada a homogeneidade da massa, ocorrência de retração da massa e ruptura de zona pelúcida. Os resultados foram expressos em porcentagem e foram submetidos ao teste do Qui-quadrado com P < 0.05. Os embriões vitrificados na presença de sacarose apresentaram menores proporções de embriões de menor qualidade após a descongelação (22,20 vs. 44,50%), e maiores percentuais de embriões homogêneos após a descongelação (63,89 vs. 38,89%) enquanto com relação aos demais parâmetros não existiram diferenças entre grupos. Pode-se concluir que a adição de 0,4 M de sacarose durante os procedimentos de vitrificação e descongelação beneficia a qualidade ...

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Section: Resultssupporting

confidence: 93%

Section: Vitrification and Warmingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

et al. 2015

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“…Embryos produced in vitro or manipulated in some manner (e.g., biopsied or bisected) usually yield lower pregnancy rates. Under optimal conditions, pregnancy rates obtained after transfer of frozen-thawed in vivo derived embryos is typically 60-70% in cattle 6,7 , 65-75% in sheep 8,9,10,11 , and 60-70% in goats 12,13,14,15 . Similarly, pregnancy rates obtained after transfer of frozen-thawed in vitro produced embryos is typically 40-50% in cattle 6,16 , 25-35% in sheep 17 , and 30-40% in goats 18 .…”

Section: Representative Resultsmentioning

confidence: 99%

Cryopreservation of Preimplantation Embryos of Cattle, Sheep, and Goats

Youngs¹

2011

JoVE

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Preimplantation embryos from cattle, sheep, and goats may be cryopreserved for short-or long-term storage. Preimplantation embryos consist predominantly of water, and the avoidance of intracellular ice crystal formation during the cryopreservation process is of paramount importance to maintain embryo viability. Embryos are placed into a hypertonic solution (1.4 -1.5 M) of a cryoprotective agent (CPA) such as ethylene glycol (EG) or glycerol (GLYC) to create an osmotic gradient that facilitates cellular dehydration. After embryos reach osmotic equilibrium in the CPA solution, they are individually loaded in the hypertonic CPA solution into 0.25 ml plastic straws for freezing. Embryos are placed into a controlled rate freezer at a temperature of -6°C. Ice crystal formation is induced in the CPA solution surrounding the embryo, and crystallization causes an increase in the concentration of CPA outside of the embryo, causing further cellular dehydration. Embryos are cooled at a rate of 0.5°C/min, enabling further dehydration, to a temperature of -34°C before being plunged into liquid nitrogen (-196°C). Cryopreserved embryos must be thawed prior to transfer to a recipient (surrogate) female. Straws containing the embryos are removed from the liquid nitrogen dewar, held in room temperature air for 3 to 5 sec, and placed into a 37°C water bath for 25 to 30 sec. Embryos cryopreserved in GLYC are placed into a 1 M solution of sucrose for 10 min for removal of the CPA before transfer to a recipient (surrogate) female. Embryos cryopreserved in EG, however, may be directly transferred to the uterus of a recipient.

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“…However, other cryoprotectants such as acetamide, glycerol, raffinose, and dimethylsulfoxide (DMSO) can be used in several combinations. Ethylene glycol and glycerol [146,170] or ethylene glycol and DMSO [8,18,50,171] are often employed. Varago et al [172] used dimethylformamide and ethylene glycol to vitrify ovine embryos but concluded that the conventional freezing with ethylene glycol was the most efficient method to cryopreserve ovine embryos.…”

Section: Cryoprotectantsmentioning

confidence: 99%

Cryopreservation of Sheep Produced Embryos – Current and Future Perspectives

Romão¹,

Marques²,

Bettencourt³

et al. 2016

Insights From Animal Reproduction

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Due to economical and scientific limitations, sheep embryo reproductive technologies are less commercially applied than in other animal species. However, it is very clear that, in the near future, those techniques are expected to have a central role in animal production as a consequence of genetic and reproductive demands. One drawback is that results obtained after sheep embryo cryopreservation are unattractive for commercial purposes. It is expected that a successful cryopreservation of sheep embryos can push forward all other reproductive biotechnologies in this species, such as multiple ovulation and embryo transfer (MOET), artificial insemination, or in vitro production of embryos. This paper tries to discuss the current and future perspectives of cryopreservation of in vivo-and in vitro-produced sheep embryos concerning advantages and limitations for its practical use and possible solutions for improving methods to allow a higher survival rate of cryopreserved embryos.

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Improved vitrification method allowing direct transfer of goat embryos

Cited by 45 publications

References 41 publications

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

Cryopreservation of Preimplantation Embryos of Cattle, Sheep, and Goats

Cryopreservation of Sheep Produced Embryos – Current and Future Perspectives

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