Successful vitrification of early-stage porcine cloned embryos |
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| Institution: | 1. Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;2. Doctorado en Ciencias Biológicas y de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;3. Departamento de Biología de la Reproducción, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;4. Departamento de Medicina y Cirugía Animal, Universidad de Murcia, Espinardo 30100, Spain;1. Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, PR China;2. Isotope Research Laboratory, College of Life Science, Sichuan Agricultural University, Ya''an 625014, PR China;3. Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, UT, USA;1. Veterinary Reproduction and Obstetrics, Department of Large Animal Sciences, University of Copenhagen, DK1870 Frederiksberg C, Denmark;2. Department of Animal Science, Aarhus University, DK8830 Tjele, Denmark;3. Institute for Farm Animal Genetics (FLI), Neustadt, Germany;4. Department of Biomedicine, Aarhus University, DK8000 Aarhus C, Denmark;5. Department of Clinical Medicine, Aarhus University, DK8200 Aarhus N, Denmark;6. Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, DK1870 Frederiksberg C, Denmark;1. College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu 210095, China;2. Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China;3. Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Shanghai 201106, China |
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| Abstract: | The objective of this study was to investigate the survival and development of porcine cloned embryos vitrified by Cryotop carrier at the zygote, 2- and 4-cell stages. The quality of resultant blastocysts was evaluated according to their total cell number, apoptotic cell rate, reactive oxygen species (ROS) production, glutathione (GSH) content and mRNA expression levels of genes related to embryonic development. The survival rates of zygotes, 2- and 4-cell embryos after vitrification did not differ from those of their fresh counterparts. Vitrification still resulted in significantly decreased blastocyst formation rates of these early-stage embryos. Moreover, the total cells, apoptotic rate, ROS and GSH levels in resultant blastocysts were unaffected by vitrification. The mRNA expression levels of PCNA, CPT1, POU5F1 and DNMT3B in the blastocysts derived from vitrified early-stage embryos were significantly higher than those in the fresh blastocysts, but there was no change in expression of CDX2 and DNMT3A genes. In conclusion, our data demonstrate that the early-stage porcine cloned embryos including zygotes, 2- and 4-cells can be successfully vitrified, with respectable blastocyst yield and quality. |
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| Keywords: | Pig Cloned embryo Vitrification Early-stage Development |
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