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Effects of different culture media, temperatures and cooling times on the development and quality of in vitro-derived bovine embryos

Grant number: 13/15905-9
Support type:Scholarships in Brazil - Master
Effective date (Start): January 01, 2014
Effective date (End): September 30, 2015
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Eunice Oba
Grantee:Fernanda Nunes Marqui
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

This study will be carry out to investigate the effects of different culture media, temperatures and cooling times on the development and quality of in vitro -derived bovine embryos. The following experiments will be performed: exp. 1, embryos cultured in m-SOF medium at temperatures of 5°C (for 24 and 48 h) and 25°C (24 h; control group); exp. 2, embryos cultured in BotuEmbryo® medium at temperatures of 5°C (for 24 e 48 h), 25°C (24 h), and the best result of exp. I; exp. 3, embryos cultured in Syngro® medium at temperatures of 5°C (for 24 e 48 h), 25°C (24 h), and the best result of exp. II. Oocytes obtained from slaughtered cows' ovaries will be selected in base maturation medium b-MM. Groups of 20-25 oocytes/drop will be matured in b-MM with additions of FSH and LH. Sperm selection will be performed in discontinuous Percoll gradient; IVF will be performed in TALP medium plus PHE and heparin. Embryo culture will be in modified SOF medium; humidified atmosphere with 5% CO2, at 38.7°C in air will be used for IVM, IVF, and IVC steps. On day-7 post-insemination blastocysts will be randomly assigned to the treatment groups. Rates of blastocyst expansion (24 h) and hatching (48h), as well as the Total Cell Number (TCN) and gene expression (genes SOD1, AOP-1, Hsp70.1 e survivin) will be evaluated after the given cooling times. ANOVA and Bonferroni T test will be used to determine differences among groups for embryo development, TCN and percentage of dead and live cells. The results of gene expression will be analyzed by x2 test. Differences of P<0.05 will be taken as significant. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MARQUI, FERNANDA N.; MARTINS, JR., ALICIO; CARDOSO, TEREZA C.; CRUZ, TAIRINI E.; DELL'AQUA, JR., JOSE ANTONIO; OBA, EUNICE. Cooling of in vitro-produced bovine embryos: Effects of medium and time on gene expression, DNA fragmentation and embryonic survival. LIVESTOCK SCIENCE, v. 230, DEC 2019. Web of Science Citations: 0.
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
MARQUI, Fernanda Nunes. Efeitos de diferentes meios e tempos de refrigeração sobre a viabilidade gênica de embriões produzidos in vitro. 2015. Master's Dissertation - Universidade Estadual Paulista "Júlio de Mesquita Filho" Faculdade de Medicina Veterinária e Zootecnia..

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