In vitro gamete derivation and production from porcine induced pluripotent stem cells (piPSCs)

Abstract

Germ cells are a source of totipotence, reproduction, inheritance, and evolution. Oocytes are a unique type of cell that gives the nucleus totipotency after fertilization; these cells can generate a new individual and contribute to genetic diversity. For this reason, many efforts have been made to clarify the fundamental mechanisms of the development of germ cells; and a wide range of in vitro models has been tested, mainly in humans and mice. One of the most promising models for in vitro reconstitution of oogenesis is pluripotent stem cells, which eventually produce functional oocytes. This technology has made significant advances in recent years, mainly in mouse models and recently in humans. However, in other species of domestic animals such as pigs, it has never been reported. In this context, this study aims, unprecedentedly, to a porcine translational model capable of producing mature germ cells in vitro from porcine induced pluripotent stem cells (piPSCs). For this, piPSCs will be initially differentiated into primordial germ cells (pPGCLCs) and then in mature gametes using the rOvary culture system described in humans. All cells will be analyzed for morphology, gene expression, protein, and epigenetic modification during different cultivation periods. The results generated in this proposal may, in an unprecedently, contribute to the generation of an advantageous model that allows the understanding of the fundamental processes of gametogenesis in porcine and other domestic animals. Also, it allows the application of an alternative source of oocytes for reproduction in animals since applicability in the production of porcine oocytes in vitro is low. The success of this technique will allow an improvement in the areas of animal reproduction, transgenics, cloning or xenotransplantation, and Precision Agriculture, mainly in the reduction of the interval between generations, acquiring a genetically superior herd faster, and in production of disease-resistant animals. Finally, the objective is to evaluate the efficacy of the porcine model in future studies in biotechnology area related to germ cell defects or other diseases that cause infertility or lack of functional gametes. (AU)