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Study of the pluripotency potential of equine stem cells derived from adult tissue and umbilical cord, submitted to genetically induced reprogramming (IPS cells)

Grant number: 13/17955-3
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): August 01, 2014
Effective date (End): December 31, 2016
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Flávio Vieira Meirelles
Grantee:Laís Vicari de Figueiredo Pessôa
Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

It has been proven that insertion of embryonic transcription factors on adult somatic cells is capable of reprogramming those cells into a pluripotent state turning them in induced pluripotent stem cells (iPS). This method has raised fundamental questions regarding the influence of the origin of the cell used to produce iPS cells. Once large mammals, like equine, are good models for human studding human diseases, interest in research involving these animals is increasing and the practice of cell therapy using mesenchymal cells on orthopedic and joint injuries is frequently in horses. Hypothesizing equine mesenchymal cells obtained from different tissues in vivo present variable pluripotent characteristic and in vitro differentiation capability, due to their derivation source; this proposal aims to generate and analyze equine iPS cells obtained by direct reprogramming of equine adult fibroblasts and mesenchymal cells derived of bone marrow, umbilical cord tissue and umbilical cord blood, regarding their in vitro pluripotency maintenance potential through endogenous Oct4 expression, marked by endogenous Oct4 promoter labeled with GFP expression and through capacity of differentiation into therapeutic interesting cell populations. iPS cells will be produced through viral transduction of human OKSM transduction factors from equine mesenchymal cells derived of bone marrow, umbilical cord tissue and umbilical cord blood. These cells will be characterized by analyses regarding cell morphology, genic expression of Oct4, Sox2 and Nanog through Real Time PCR, alkaline phosphatase expression and in vitro and in vivo differentiation into cell populations of interest. As expected results, we intent to observe low or none endogenous Oct4 expression on mesenchymal stem cells; expression of exogenous pluripotent related factors between 4-6days after transduction; endogenous expression of factors related to pluripotency after 14 days of reprogramming; in vivo teratoma formation containing at least one cell type of each germ layer; iPS cells derived from mesenchymal cells that present higher multipotent potencial must present higher pluripotenciality; after iPS differentiation into cell populations of interest, these cells mustn't show endogenous Oct4 expression, a characteristic of full reprogramming. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DE FIGUEIREDO PESSOA, LAIS VICARI; LISBOA PIRES, PEDRO RATTO; DEL COLLADO, MAITE; GODOY PIERI, NAIRA CAROLINE; RECCHIA, KAIANA; SOUZA, ALINE FERNANDA; PERECIN, FELIPE; DA SILVEIRA, JULIANO COELHO; CESAR DE ANDRADE, ANDRE FURUGEN; AMBROSIO, CARLOS EDUARDO; et al. Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues. STEM CELLS INTERNATIONAL, . (08/57877-3, 13/17955-3, 15/26818-5, 13/08135-2, 12/50533-2, 14/22887-0)
DE FIGUEIREDO PESSOA, LAIS VICARI; LISBOA PIRES, PEDRO RATTO; DEL COLLADO, MAITE; GODOY PIERI, NAIRA CAROLINE; RECCHIA, KAIANA; SOUZA, ALINE FERNANDA; PERECIN, FELIPE; DA SILVEIRA, JULIANO COELHO; CESAR DE ANDRADE, ANDRE FURUGEN; AMBROSIO, CARLOS EDUARDO; et al. Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues. STEM CELLS INTERNATIONAL, v. 2019, p. 15-pg., . (14/22887-0, 08/57877-3, 12/50533-2, 15/26818-5, 13/17955-3, 13/08135-2)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
PESSÔA, Laís Vicari de Figueiredo. Pluripotency potential of equine mesenchymal cells obtained from different sources subjected to genetically induced reprogramming (iPS cells). 2016. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD) São Paulo.

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