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Influence of embryo culture medium (BARC) supplementeded with different macromolecules on in vitro bovine oocyte maturation

Grant number: 11/11985-2
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2011
Effective date (End): January 31, 2013
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Alicio Martins Júnior
Grantee:Jéssica de Oliveira Caldeira
Home Institution: Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil
Associated research project(s): 12/20723-4

Abstract

This study will be carry out to assess the influence of bovine embryo culture medium (BARC), supplemented with BSA, CFS or PVA, on the in vitro oocyte maturation, evidenced by embryo production and quality at expanded blastocyst stage. Four experiments will be performed, as follows: exp.1: addition of FCS to BARC medium at concentrations of 0, 5 and 10%; exp. 2: addition of BSA to BARC medium at concentrations of 0, 4 and 8 mg/ml; exp. 3: addition of PVA to BARC medium at concentrations of 0, 0.5 and 1.0 mg/ml; exp. 4: comparisons among maturation media supplemented with FCS, BSA or PVA, with evaluations of embryonic development up to mentioned stages, total cell number (TCN) and apoptosis. Oocytes obtained from cow ovaries at slaughterhouse will be selected (PBS + FCS, BSA or PVA) and matured (groups of 20 oocytes/drop/100mL) in BARC medium supplemented with gonadotropins and penicillin, according to the experimental design. Percoll gradient will be used for sperm selection and TALP medium for IVF. In vitro embryo culture will be performed in SOF-m medium; a humidified atmosphere with 5% CO2, in air, at 38.7oC will be used for all steps. The number of oocytes reaching morula, blastocyst and expanded blastocyst stages will be recorded, respectively at 72, 144 and 168 h post-insemination. ANOVA and Bonferroni t test will be used to determine differences among groups (embryo development). The data for apoptosis and TCN will be analyzed by Mann-Whitney e one -way repeated measures ANOVA, respectively. Differences of p<0.05 will be taken as significant.