Effects of different media and cooling times on viability and gene expression of in vitro-derived bovine embryos

Author(s)

Fernanda Nunes Marqui

• Document type: Master's Dissertation
• Press: Botucatu. 2018-07-27.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Medicina Veterinária e Zootecnia. Botucatu
• Defense date: 2015-09-30
• Advisor: Eunice Oba; Alicio Martins Júnior

Abstract

This study was carried out to investigate the effects of different media and cooling times on the development and gene expression of in vitro-derived bovine embryos. Therefore, blastocysts D7 were divided into three groups: control, embryos kept in SOFaa medium, under CO2 incubation, for 6, 24 and 48 hours; cooled embryos, kept in Medium 199, for 24 (M199-24h) and 48 hours (M199-48h) or in BotuEmbryo® medium, for 24 (BE-24h) and 48 hours (M199-48h). After cooling, the embryos were cultured under the same conditions used for the control group for evaluation of embryo viability at 6 and 48 hours, and of re-expansion and hatching rates, at 24 and 48 hours, respectively. At 6h of culture part of the embryos was removed and stored for later analysis of qPCR and TUNEL. Low percentage (P<0.05) of viable blastocysts, at 6 and 48 hours, was observed in group M199-48h than for control group. Re-expansion rate did not differ between cooled embryos and control group, however, lower (P<0.05) hatching rate was observed in M199-48h and BE-48h groups than for control group. Higher (P<0.05) percentage of apoptotic cells was found in BE-48h compared with control and BE-24h groups. Media and cooling times used did not alter the expression of genes HSPA1A, PRDX1, SOD1 and SLC2A1. Thus, overall, medium and cooling time did not promoted potential adverse effect on the embryonic viability, re-expansion and hatching rate, number of apoptotic cells and gene expression, suggesting the use of BotuEmbryo® medium, which is commercially available (AU)

• FAPESP's process: 13/15905-9 - Effects of different culture media, temperatures and cooling times on the development and quality of in vitro-derived bovine embryos
• Grantee: Fernanda Nunes Marqui
• Support Opportunities: Scholarships in Brazil - Master