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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues

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Author(s):
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de Figueiredo Pessoa, Lais Vicari [1, 2] ; Lisboa Pires, Pedro Ratto [1] ; del Collado, Maite [1] ; Godoy Pieri, Naira Caroline [3] ; Recchia, Kaiana [1] ; Souza, Aline Fernanda [1] ; Perecin, Felipe [1] ; da Silveira, Juliano Coelho [1] ; Cesar de Andrade, Andre Furugen [3] ; Ambrosio, Carlos Eduardo [1] ; Bressan, Fabiana Fernandes [1] ; Meirelles, Flavio Vieira [1]
Total Authors: 12
Affiliation:
[1] Univ Sao Paulo, Dept Med Vet, Fac Zootecnia & Engn Alimentos, BR-13635000 Pirassununga - Brazil
[2] Univ Copenhagen, Dept Vet & Anim Sci, Sect Anat & Biochem, DK-1870 Frederiksberg C - Denmark
[3] Univ Sao Paulo, Dept Reprod Anim, Fac Med Vet & Zootecnia, BR-13635000 Pirassununga - Brazil
Total Affiliations: 3
Document type: Journal article
Source: STEM CELLS INTERNATIONAL; 2019.
Web of Science Citations: 1
Abstract

Introduction. Pluripotent stem cells are believed to have greater clinical potential than mesenchymal stem cells due to their ability to differentiate into almost any cell type of an organism, and since 2006, the generation of patient-specific induced pluripotent stem cells (iPSCs) has become possible in multiple species. Objectives. We hypothesize that different cell types respond differently to the reprogramming process; thus, the goals of this study were to isolate and characterize equine adult and fetal cells and induce these cells to pluripotency for future regenerative and translational purposes. Methods. Adult equine fibroblasts (eFibros) and mesenchymal cells derived from the bone marrow (eBMmsc), adipose tissue (eADmsc), and umbilical cord tissue (eUCmsc) were isolated, their multipotency was characterized, and the cells were induced in vitro into pluripotency (eiPSCs). eiPSCs were generated through a lentiviral system using the factors OCT4, SOX2, c-MYC, and KLF4. The morphology and in vitro pluripotency maintenance potential (alkaline phosphatase detection, embryoid body formation, in vitro spontaneous differentiation, and expression of pluripotency markers) of the eiPSCs were characterized. Additionally, a miRNA profile analysis of the mesenchymal and eiPSCs was performed. Results. Multipotent cells were successfully isolated, but the eBMmsc failed to generate eiPSCs. The eADmsc-, eUCmsc-, and eFibros-derived iPSCs were positive for alkaline phosphatase, OCT4 and NANOG, were exclusively dependent on bFGF, and formed embryoid bodies. The miRNA profile revealed a segregated pattern between the eiPSCs and multipotent controls: the levels of miR-302/367 and the miR-92 family were increased in the eiPSCs, while the levels of miR-23, miR-27, and miR-30, as well as the let-7 family were increased in the nonpluripotent cells. Conclusions. We were able to generate bFGF-dependent iPSCs from eADmsc, eUCmsc, and eFibros with human OSKM, and the miRNA profile revealed that clonal lines may respond differently to the reprogramming process. (AU)

FAPESP's process: 12/50533-2 - GIFT: genomic improvement of fertilization traits in Danish and Brazilian cattle
Grantee:Marcelo Fábio Gouveia Nogueira
Support type: Research Projects - Thematic Grants
FAPESP's process: 13/17955-3 - Study of the pluripotency potential of equine stem cells derived from adult tissue and umbilical cord, submitted to genetically induced reprogramming (IPS cells)
Grantee:Laís Vicari de Figueiredo Pessôa
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 08/57877-3 - National Institute of Science and Technology in Cell Therapy
Grantee:Roberto Passetto Falcão
Support type: Research Projects - Thematic Grants
FAPESP's process: 15/26818-5 - Investigation of cellular and molecular mechanisms on in vitro induced toti- and pluripotency acquisition - a translational approach
Grantee:Fabiana Fernandes Bressan
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 14/22887-0 - Cell-secreted vesicles containing miRNAs modulate epigenetic changes during in vitro culture of bovine gametes and embryos
Grantee:Juliano Coelho da Silveira
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
Grantee:Dimas Tadeu Covas
Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC