Bovine in vitro oocyte maturation and embryo culture in liquid marbles 3D culture system

Author(s)

Ferronato, Giuliana de Avila ; dos Santos, Carolina Monica ; Rosa, Paola Maria da S. ; Bridi, Alessandra ; Perecin, Felipe ; Meirelles, Flavio Vieira ; Sangalli, Juliano Rodrigues ; da Silveira, Juliano Coelho

• Document type: Journal article
• Source: PLoS One; v. 18, n. 4, p. 17-pg., 2023-04-21.

Abstract

Despite the advances in in vitro embryo production (IVP) over the years, the technique still has limitations that need to be overcome. In cell cultures, it is already well established that three-dimensional culture techniques are more physiological and similar to the in vivo development. Liquid marble (LM) is a three-dimensional system based on the use of a hydrophobic substance to create in vitro microbioreactors. Thus, we hypothesized that the LM system improves bovine in vitro oocyte maturation and embryo culture. In experiment I, bovine cumulus-oocyte complexes (COCs) were placed for in vitro maturation for 22h in two different groups: control (conventional 2D culture) and LM (three-dimensional culture). We found that oocyte nuclear maturation was not altered by the LM system, however it was observed a decrease in expression of genes important in the oocyte maturation process in cumulus cells of LM group (BCL2, EIF4E, and GAPDH). In experiment II, the COCs were conventionally matured and fertilized, and for culture, they were divided into LM or control groups. There was a decrease in blastocyst rate and cell counting, a down-regulation of miR-615 expression, and an increase in the DNA global methylation and hydroxymethylation in embryos of LM group. Therefore, for the bovine in vitro embryo production, this specific three-dimensional system did not present the advantages that we expected, but demonstrated that the embryos changed their development and epigenetics according to the culture system. (AU)

• FAPESP's process: 19/25675-7 - In vitro production of bovine embryos in different 3D culture systems: effects on nuclear morphology, methylation pattern, gene transcription and extracellular vesicles
• Grantee: Giuliana de Avila Ferronato
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 14/22887-0 - Cell-secreted vesicles containing miRNAs modulate epigenetic changes during in vitro culture of bovine gametes and embryos
• Grantee: Juliano Coelho da Silveira
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 21/06645-0 - Extracellular vesicles as a platform for diagnostic and manipulation of the in vitro embryo production system: the next generation in animal reproduction
• Grantee: Juliano Coelho da Silveira
• Support Opportunities: Research Grants - Young Investigators Grants - Phase 2