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Induced reprogramming of bovine somatic cells into induced pluripotent cells (IPS) and primordial germ cells-like (PGCLs)

Grant number: 13/13686-8
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): October 01, 2013
Effective date (End): September 30, 2016
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Flávio Vieira Meirelles
Grantee:Fabiana Fernandes Bressan
Home Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

Recent technologies, for example nuclear transfer and genetic induced reprogramming, have been employed to induce somatic cells into a pluripotent state similar to embryonic status. The process of nuclear reprogramming is highly desirable and presents important contributions for both basic and applied science for example, increasing the efficiency of reproductive biotechnologies aiming animal production or regenerative medicine. Several studies have reported that induced pluripotente stem cells (iPSCs), similarly to embryonic stem cells (ES cells), are able to differentiate and give rise to primordial germinal stem cells (PGCs) and are even able to develop into gametes (oocytes and spermatozoa). Regarding regenerative medicine, the in vitro production of PGCs from iPS cells represents and important tool for reproductive biotechnologies, providing an ideal platform for its potential use on diagnosis and treatment of several infertilities, as well as elucidating the molecular mechanisms overlying PGCs development and biotechnologies related to animal production.The efficiency of reprogramming into pluripotency, as well as its subsequent differentiation into PGCs-like cells (PGCCLS) still presents low efficiency, hampering its use in translational and regenerative medicine. A probable explanation is the failure on epigenetic reprogramming leading to an incomplete restore of the pluripotent cellular status, especially, during the acquisition and maintenance of genomic imprintings periods.Therefore, the present proposal aims the generation of germinal cells trough reprogramming of somatic cells induced to pluripotency, and also to characterize temporal profiles of gene expression and epigenetics during the process of dedifferentiation and cellular differentiation in the bovine model, allowing the study of allele specific gene expression of imprinted genes related to early development. Hence, global gene expression and epigenetics of cells submitted to in vitro induction of pluripotency and subsequent differentiation into PGCCLs. Results obtained from this proposal will provide important and still unreported insights regarding molecular mechanisms involved in nuclear reprogramming.