Evaluation of lipid drops in the in vitro maturation of ovarian with low and high count of antral follicles

• Grant number: 18/09006-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2018
• End date: June 30, 2020
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Sheila Merlo Garcia Firetti
• Grantee: Leandro Francisco da Silva
• Host Institution: Faculdade de Ciências Agrárias. Universidade do Oeste Paulista (UNOESTE). Campus de Presidente Prudente. Presidente Prudente , SP, Brazil

Abstract

The objective of this project is to evaluate the quality of in vitro maturation, according to the amount of lipid droplets, of oocytes from slaughterhouse ovaries, with low or high antral follicle count (AFC) and to identify the group that presents greater viability for the in vitro fertilization procedure. Five replicates will be performed, and in each repetition the ovaries will be separated into two groups: high AFC and low AFC. The visual AFC should include all visible follicles, e 3 millimeters in diameter and the numerical limits will be defined according to the distribution of data by performing statistical tests. After separation of the groups the visible follicles will be aspirated, and the selected oocytes, classified according to the morphological aspect of the CCO's, and taken separately (according to the group and degree of classification), for maturation for 24h, 39°C, and 5% CO2 in air. After maturation, they were stained with Sudan Black B 1% (199664, Sigma Co., St. Louis, USA), and evaluated individually under a light microscope (DMLS; Leica), to evaluate the amount of lipid droplets in each oocyte . Statistical analysis of the data will be performed by non-parametric tests, with the effect of the presence of low or high scores verified using the Chi-square test (Ç2) at a significance level of 5% and multiple comparisons using Fisher's exact test , using statistical software "R" (R core Team, 2013).