Generation of induced pluripotency stem cells (iPSCs) from adult cells of small red deer (Mazama bororo)

• Grant number: 20/08562-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2020
• End date: March 31, 2022
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Fabiana Fernandes Bressan
• Grantee: Luana Scarpelini Rosa
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Associated research grant: 15/26818-5 - Investigation of cellular and molecular mechanisms on in vitro induced toti- and pluripotency acquisition - a translational approach, AP.JP

Abstract

Brazil is one of the countries with the richest biodiversity in the world, however it is constantly threatened by the negative consequences inflicted by human activity on the environment. Currently, 1173 Brazilian animal species are considered threatened, one of them being the small red brocket (Mazama bororo), an animal endemic to the Atlantic Forest biome, believed to have the smallest geographic distribution of all the cervidae family. Mazama bororo is considered vulnerable by the IUCN and by Brazil's environmental ministry. Several institutions allied with public policies have sought conservation measures to help preserve these species found at risk of extinction. Conservation programs include actions that go from protecting the animals' habitat to preserving their genetic material, the last being essential for maintaining the animals' resilience and resistance. In this context, reproductive biotechnologies prove to be very important for preserving animals' genetic material. In this study, we aim to generate and characterize induced pluripotent stem cells(iPSCs) cells originated from adult fibroblast belonging to Mazama bororo. For that, fibroblasts previously collected and stored will be cultured and transduced with polycistronic lentiviral vectors that express human or murine OCT4, SOX2, KLF4 and c-MYC (hOSKM or mOSKM). The generated iPSCs will be characterized by alkaline phosphatase detection, immunocytochemistry for OCT4, SOX2 and NANOG pluripotency proteins, formation of embryoid bodies differentiation in vitro, gene expression analysis and teratoma assay. The generation of iPSCs from somatic cells is very promising in human and veterinary medicine, and possibilities for their use ranges from studies of diseases and regenerative therapies to possible correction of infertilities, including the generation of gametes and embryos in invitro from previously stored cells and genetic material banks. To date, there is no knowledge of generation of iPSC cells from the cervid family in the literature. Obtaining the iPSCproposed in this study will be a major contribution to the reproductive biotechnologies used in the conservation of the species itself and also to other animals of the cervidae family.