In vitro embryo generation from induced pluripotent stem cells in cattle

• Grant number: 23/11635-9
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2024
• End date: March 31, 2025
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Fabiana Fernandes Bressan
• Grantee: Raiane Cristina Fratini de Castro
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

In vitro cultivation of embryos facilitates the understanding of formation mechanisms and developmental stages usually inaccessible during early embryonic development. Induced pluripotent stem cells (iPSCs) are a useful tool to facilitate the application of advanced reproductive technologies, contributing to the generation of animals of high genetic merit in a short period of time. Thus, the goal of the research will be to perform the in vitro production of embryos derived from bovine induced pluripotent stem cells (biPSCs) through complementation with cells from the trophectoderm, forming an embryonic aggregation morphologically similar to a blastocyst. In vitro embryo production and differentiation of biPSCs in culture medium supplemented with BMP4, A83-01 and PD173074 will be performed in trofectoderm cells. Subsequently, experimental groups will be formed and cultured in a non-adherent suspension shaking system. Characterization of the embryonic structures produced will occur through immunocytochemistry testing of EOMES, OCT4 and CDX2 genes expressed during embryonic development, morphological evaluation and quantitative RT-PCR blastocyst marker genes GATA3, SOX2, CDX2, SOX17 and pluripotency marker genes NANOG and OCT4. It is expected to define an in vitro model that leads to the formation of embryos capable of recapitulating the general morphology of the blastocyst from biPSCs supplemented with cells from the trophectoderm, aiming to provide a methodology that can serve as a model for tests with other species. The aim is to contribute to the improvement of reproductive efficiency within veterinary medicine, as well as to the translational study of early mammalian embryonic development.