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Generation of induced pluripotency stem cells (iPSCs) from adult camelid cells

Grant number: 20/07921-8
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): September 01, 2021
Effective date (End): August 31, 2022
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal researcher:Fabiana Fernandes Bressan
Grantee:Nayanne Sant Clair Cardoso da Silva
Home 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

Representatives of the family Camelidae infraorder Tylopoda, such as alpacas, guanacos, vicunas, llamas, camels, dromedaries, amongst others, have a evident presence in four (Americas, Asia, Africa, and Australia) from the seven continents and also have great economic and social importance due to the ability to adapt to adverse conditions, typical of the arid regions they inhabit, their ability to submit to numerous tasks, such as transportation, agricultural work, leisure, production of wool, meat, milk and their derivatives. The present study has as main objective to generate a greater knowledge about the developmental biology and new technologies in these animals, and in particular, aiming in the future to contribute to the preservation of the species, considering the threat of extinction of some camelids in the wild, like vicuñas (almost wiped out by poachers) and camels. To this end, due to the role of stem cells to enable reproductive efficiency and the possibility of innovative therapies, in this study we sought, in an unprecedented way, the production and characterization of induced pluripotent stem cells (iPSC) through in vitro reprogramming of in vitro adult cells into a pluripotent state, similar to the embryonic, in camelids. For that, fibroblasts derived from Vicugna pacos obtained through biopsies will be isolated, cultured and transduced with polycistronic lentiviral vectors that express human or mouse OCT4, SOX2, KLF4 and cMYC (hOSKM or mOSKM). iPSCs will be analyzed regarding detection of alkaline phosphatase and immunocytochemistry for OCT4, SOX2 and NANOG pluripotency proteins, embryoid body formation and differentiation in vitro, gene expression analysis and teratoma assay. The present proposal is relevant both as a model for studying diseases, developing new drugs and countless therapeutic actions and innovations; and for promoting advanced reproductive technologies, such as the possibility of generating embryos and gametes in vitro, playing an important role in preservation of the species.

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