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In vitro production of bovine embryos in different 3D culture systems: effects on nuclear morphology, methylation pattern, gene transcription and extracellular vesicles

Grant number: 19/25675-7
Support type:Scholarships in Brazil - Master
Effective date (Start): June 01, 2020
Effective date (End): February 28, 2022
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Juliano Coelho da Silveira
Grantee:Giuliana de Avila Ferronato
Home Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
Associated research grant:14/22887-0 - Cell-secreted vesicles containing miRNAs modulate epigenetic changes during in vitro culture of bovine gametes and embryos, AP.JP

Abstract

In vitro embryo production (IVP) is a technique of high relevance and use, however it still has some limitations that end up reducing its efficiency, such as low development rate to the blastocyst stage and low pregnancy rates. Embryos obtained from IVP still differ from embryos produced in vivo in epigenetic factors, which can interfere with the establishment of pregnancy and also the health of animals after birth. In somatic cell cultivation there is a well established alternative for improving the technique which is 3D cultivation. Thus, it is hypothesized that it is possible to improve the IVP technique using a 3D culture system in order to increase the initial embryonic developmental rate of IVP and decrease the epigenetic differences that exist between embryos produced in vivo and in vitro. Thus, the objective of this project is to evaluate two different 3D cultivation systems, alginate and magnetic nanoparticle matrices, for IVP, through embryonic quality analysis, cellular communication by extracellular vesicle profile (EVs) and methylation pattern. global DNA hydroxymethylation. For this, the project will have three experiments that will be conducted after the fertilization stage, with the following embryo cultivation groups: experiment 1 - control group and alginate matrix group; experiment 2 - control group and magnetic nanoparticle group and; Experiment 3 - In vitro control group, alginate matrix group, nanoparticle group and in vivo embryo production group. (AU)