Improvement in the production of bovine tetraploid embryos through the fusion of two cell zone-free

FELIPE, MINERVA YAURI; DANIEL F. SALAMONE

Bologna

Congreso; 19th ICAR International Congress of Animal Reproduction; 2023

ICAR

IMPROVEMENT IN THE PRODUCTION OF BOVINE TETRAPLOID EMBRYOS THROUGH THE FUSION OF TW0-CELL ZONE-FREE EMBRYOM Yauri Felipe1 D Salamone11. Laboratorio de Biotecnología Animal, FAUBA/INPA-CONICET, Argentina.Tetraploid embryos differentiate into trophectoderm when they are complemented with diploid embryos. The objective of this study was to optimize the electrofusion of two-cell embryos to generate tetraploid (4n) blastomeres in the presence or absence of zona pellucida (ZP) at two different voltages. After 29 up to 30 hours of initiating the IVF, all embryos with two blastomeres were randomly assigned to the following groups a) ZP free fused with 0.8 Kv/cm (ZP-Free-0.8), b) ZP free fused with 1 Kv/cm (ZP-Free-1), b) intact ZP with 0.8 Kv/cm (iZP-0.8), c) intact ZP with 1 Kv / cm (iZP-1). For ZP-free embryos, ZP was removed by a treatment with a protease. Then embryos were treated with 0.8 Kv/cm or 1 Kv/cm for the fusion of the two cells of each embryo. Each treatment involved 2 pulses of 30 μs separated by 0.1s of rest. Two hours after fusion, presumptive 4n zygotes were cultured independently in microwells with synthetic oviductal fluid (SOF) medium in 6.5% CO2 in air at 38.5°C for 7 days. The best fusion rate was observed in the ZP-Free-0.8 group, with 74.83% of fused embryos, compared to 55.75%; 60.37%, and 38.78% for the groups ZP-Free-1; iZP-0.8 and iZP-1 respectively (p