Assessment of the Metabolic Profile of Bovine Embryo Co-culture with Oviductal Cell Spheroids in DNA Methylation Dynamics

• Grant number: 24/16287-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: April 01, 2025
• End date: July 31, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Marcella Pecora Milazzotto
• Grantee: Ana Beatriz dos Reis Bartoli
• Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Associated research grant: 21/11747-6 - Remote or presential: the importance of communication in the reproductive processes, AP.TEM

Abstract

Epigenetic mechanisms, such as DNA methylation and hydroxymethylation, play crucial roles in organism development, regulating the expression of genes essential for lineage determination and cellular identity. During bovine embryonic development, there is a shift in the utilization of energy substrates, which is directly linked to DNA methylation and other epigenetic mechanisms. Despite improvements in assisted reproduction technologies, in vitro production systems still fail to fully replicate physiological systems, as evidenced by disparities between in vitro and in vivo-produced embryos. Epigenetic reprogramming can also be disrupted by conditions arising from in vitro production, such as culture medium components, light exposure, temperature, and oxygen tension. Embryos subjected to in vitro culture conditions (IVC) before or during the embryonic genome activation demonstrate increased susceptibility to DNA methylation alterations. To better understand gene expression dynamics, metabolism, and in vitro embryonic development, co-culture systems employing oviductal somatic cells, such as epithelial cells, along with in vitro-produced embryos, have been developed. This approach is based on the observation that in mammals, the initial contact between the embryo and the maternal environment occurs in the oviduct, with this contact established during the first 3 to 4 days post-fertilization in cattle. Thus, the hypothesis of this project is that co-culturing embryos with oviductal cell spheroids alters the secretome of these cells, and therefore the extracellular environment, leading to a decrease in the global methylation pattern observed in early-stage in vitro embryo cultures. The project aims to determine whether the co-culture model of embryos with oviductal cell spheroids will result in alteration of the global hypermethylation pattern already observed in in vitro-produced embryos, characterizing the secretome of the co-culture medium for a better understanding of maternal-embryonic communication dynamics regarding global methylation.