A comparative analysis of the efficacy of three cryopreservation protocols on the survival of in vitro-derived cattle embryos at pronuclear and blastocyst stages |
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| Affiliation: | 1. School of Health, Medical & Applied Sciences, Central Queensland University, Rockhampton, QLD 4702, Australia;2. National Key Laboratory of Animal Cell Technology, National Institute of Animal Sciences, Hanoi, Viet Nam;3. Australian Reproductive Technologies, Mt Chalmers, QLD 4702, Australia;4. Education Program in Reproduction & Development, Department of Obstetrics & Gynaecology, Monash University, Clayton, VIC 3168, Australia;1. Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Camino Cruz Del Sur, 2250, Montevideo, Uruguay;2. Programa de Posgrado, Facultad de Veterinaria, Universidad de La República, Av. Lasplaces, 1550, Montevideo, Uruguay;3. Unidad de Animales Transgénicos y de Experimentación, Institut Pasteur de Montevideo, Mataojo, 2020, Montevideo, Uruguay;1. Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea;2. Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea;3. Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA;4. School of Veterinary Medicine, Utah State University, Logan, UT 84322-4700, USA;5. Center for Integrated Biosystems, Utah State University, Logan, UT 84322-4700, USA;1. School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil;2. Centre de Recherche en Biologie de la Reproduction, Faculté des Sciences de l’Agriculture et de l’Alimentation, Département des Sciences Animales, Pavillon INAF, Université Laval, Québec City, Quebec, Canada;3. Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, Federal District, Brazil;1. NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki 305-0901, Japan;2. Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan;3. College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea;4. Food and Fertilizer Technology Center, Taipei 10648, Taiwan |
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| Abstract: | The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrification and long equilibration vitrification) on in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages was compared. 199 expanded blastocysts of good quality were assigned randomly into four treatment groups [control, non-cryopreserved (fresh, unfrozen); and the three cryopreservation methods]. The re-expansion of the cryopreserved blastocysts after 24 h in vitro culture was similar to that of the fresh control group. However, the hatching rate of expanded blastocysts after 48 h culture was significantly less for the slow freezing group (31/47; 66.0%) than for both the short equilibration vitrification (46/51; 90.2%) and long equilibration vitrification groups (42/50; 84.0%). Denuded presumptive zygotes at the pronuclear stage (14–18 h post-insemination) were assigned randomly to the same four treatment groups and, following thawing, embryos were assessed for their capacity to cleave and to develop into a blastocyst. Overall, cleavage rates of cryopreserved zygotes were significantly less than those of the fresh control. The blastocyst formation rate of slow-frozen zygotes (4/81; 4.9%) was significantly less than that of zygotes subjected either to short equilibration vitrification (18/82; 22.0%) or long equilibration vitrification (16/74; 21.6%). All cryopreservation groups showed rates of blastocyst formation that were significantly less than that of the fresh control (51/92; 55.4%). Collectively, our findings indicate that vitrification is the preferred technology to cryopreserve in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages. Moreover, short equilibration vitrification technology can improve outcomes and be more efficient by taking less time to perform. |
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| Keywords: | Cattle Cryopreservation Pronuclear Slow freezing Vitrification |
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