Induced pluripotent cells as an alternative for viable gametes production

Abstract

The demand for animal products is increasing globally, and in vitro technologies are great allies to accelerate the process of genetic enhancement, enabling the development of animals with higher productivity. Among the biotechnologies that can be applied, induced pluripotent stem cells (iPSCs) can contribute in several ways to the acquisition of genetically superior animals or to the development of animal products in vitro. Thus, the present study proposes the noninvasive collection and in vitro isolation of cells derived from urine (urine-derived cells, UDCs) or milk (milk-derived cells, MDCs) from Holstein cows (Bos taurus). It is also proposed to in vitro reprogram UDCs and MDCs to a state of pluripotency (bovine iPSCs, biPSCs) using a non-integrative (episomal) methodology, aiming to acquire cells without exogenous DNA, it would facilitate the use of the generated biPSCs. The biPSCs will be initially differentiated inthe N2B27 culture medium supplemented with activin A, bFGF, and KSR in epiblast-like cells (EpiLCs), which will later be differentiated into primordial germ cells (PGCs) when cultured in GMEM medium supplemented with KSR, NEAA, sodium pyruvate, 2-²mercaptoethanol, Lglutamine, penicillin/streptomycin, BMP4, SCF, BMP8b and EGF. The PGCs will be specialized in viable gametes when cultured in a rOvary system, derived from murine fetal ovary cells. The results obtained in this proposal can contribute unprecedentedly to the translational study of the development of germ lines and precision agriculture, enabling the reduction of the interval between generations and faster acquisition of a genetically superior herd faster.