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Study of xygen tension influence on expression of epigenetic regulator genes in bovine fibroblasts

Grant number: 18/04638-3
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2018
Effective date (End): May 31, 2019
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Fabiana Fernandes Bressan
Grantee:Jessica Brunhara Cruz
Home Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

Artificial environment demand the best simulation to provide natural development of cell culture. Among the components of this environment, the oxygen tension is on of most important, where variations can interfere and could change the cellular epigenome. Epigenetic modifications are influenced by O2 concentration, since demethylases, enzymes responsible for removing methyl radicals from DNA and histones, are members of the Fe2+ and O2--dependent protein family by catalysis, which may influence the pattern of gene expression control. Therefore, understanding the biological mechanisms involved in this gene regulation is important to improve the quality of in vitro system to grow somatic cells. Moreover, improving this system could benefit the proper reprograming of this nuclear donor in techniques as nuclear somatic cell transfer (TNCS) and generation of induced pluripotent stem cells (IPSCs). Based on this, we hypothesized that different concentrations of oxygen during in vitro cell culture changes the transcription of genes involved on epigenetic marks regulation in bovine fibroblasts. Thus, the general objective is to analyze the expression of several genes responsible for epigenetic modification in bovine fibroblasts under different concentrations of O2. For this, bovine fibroblasts will be cultured in low (5%) and high (20%) O2 tension in different passages. At the end of each culture period, the samples will be collected for subsequent RNA extraction and regulatory genes of DNA methylation and histone modification will be analyzed. Thus, our goal is to study the effect of O2 concentration on the gene expression of epigenetic regulatory enzymes in bovine fibroblasts and, with this, assist the development of more efficient methodologies for nuclear donor cells cultures previous to nuclear reprogramming.