In vitro bovine embryo production in a synthetic medium: embryo development, cryosurvival, and establishment of pregnancy.
The aim of this study was to develop an in vitro embryo culture medium without either fetal calf serum or BSA, using various growth factors and cytokines (GFs-CYKs; IGF-I, IGF-II, bFGF, LIF, GM-CSF, TGF-β1, and PDGF-BB), and other molecules with surfactant and embryotrophic properties, such as recombinant albumin (RA) and hyaluronan (HA). The first part of the study was dedicated to define the best combination of GFs-CYKs + RA + HA for optimal embryonic development. Next, we compared development rates and embryo quality (inner cell mass [ICM]-to-total cell number [TCN] ratio), and postthaw survival and hatching rates using this synthetic medium (T1) and a control medium: synthetic oviduct fluid + BSA + ITS (insulin, transferrin, and selenium). The blastocyst rates were significantly higher with T1 than those with the control at 7 and 8 days after fertilization. There was no significant difference in TCN or the ICM/TCN ratio between the two treatments. Survival and hatching rates 48 hours after thawing were similar for both treatments. Finally, nine embryo transfers were conducted using fresh and previously frozen Day-7 blastocysts to evaluate the in vivo viability of embryos produced in this synthetic medium; four gestations were obtained from six fresh embryos and one gestation from three frozen embryos. In conclusion, the fetal calf serum and BSA-free medium, supplemented with GFs-CYKs + RA + HA, improved embryo development and gave comparable ICM/TCN ratios and postthaw survival rates to the control with BSA. Fresh and frozen embryos produced in this medium are viable for embryo transfer. This fully synthetic method of embryo culture is a useful means of reducing the risk of disease transmission via embryo transfer.
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France; Gènes Diffusion SAS, Douai, France.
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Moreno
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Neira
UMR 703, PAnTher, INRA Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Nantes, France.
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Dubreil
Gènes Diffusion SAS, Douai, France.
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Liegeois
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Destrumelle
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Michaud
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Thorin
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Briand-Amirat
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France.
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Bencharif
Laboratory of Biotechnology and Pathology of Reproduction, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Oniris, Nantes, France. Electronic address: daniel.tainturier@oniris-nantes.fr.
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Tainturier
D000418:Albumins; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology*; D015925:Cryopreservation; D003470:Culture Media / Q000737:chemistry; D016207:Cytokines; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D004624:Embryo Transfer / Q000662:veterinary; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D006820:Hyaluronic Acid; D036341:Intercellular Signaling Peptides and Proteins; D011247:Pregnancy; D011994:Recombinant Proteins; D013501:Surface-Active Agents
D000418:Albumins; D003470:Culture Media; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D006820:Hyaluronic Acid
Cytokine; Embryo culture; Growth factor; Hyaluronan; Synthetic medium