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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.)

In Vitro Induction of Pluripotency from Equine Fibroblasts in 20% or 5% Oxygen

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Author(s):
de Castro, Raquel V. G. [1, 2] ; Pieri, Naira C. G. [1, 3] ; Fantinato Neto, Paulo [1] ; Grizendi, Bianca M. [1] ; Doria, Renata G. S. [1] ; Meirelles, Flavio V. [1] ; Smith, Lawrence C. [4] ; Garcia, Joaquim M. [2] ; Bressan, Fabiana F. [1]
Total Authors: 9
Affiliation:
[1] Univ Sao Paulo FZEA USP, Fac Anim Sci & Food Engn, Dept Vet Med, BR-13635900 Sao Paulo - Brazil
[2] Sao Paulo State Univ, Fac Agr & Vet Sci, Dept Pathol Reprod & One Hlth, BR-14884900 Sao Paulo - Brazil
[3] Univ Sao Paulo FMVZ USP, Sch Vet Med & Anim Sci, Dept Anim Reprod, BR-13635900 Sao Paulo - Brazil
[4] Univ Montreal, Fac Vet Med, Ctr Rech Reprod & Fertilite, Montreal, PQ J2S 2M2 - Canada
Total Affiliations: 4
Document type: Journal article
Source: STEM CELLS INTERNATIONAL; v. 2020, NOV 26 2020.
Web of Science Citations: 0
Abstract

The cellular reprogramming into pluripotency is influenced by external and internal cellular factors, such as in vitro culture conditions (e.g., environmental oxygen concentration), and the aging process. Herein, we aimed to generate and maintain equine iPSCs (eiPSCs) derived from fibroblasts of a horse older than 20 years and to evaluate the effect of different levels of oxygen tension (atmospheric 20% O-2, 5% O-2, or 20% to 5% O-2) on these cells. Fibroblasts were reprogrammed, and putative eiPSCs were positive for positive alkaline phosphatase detection; they were positive for pluripotency-related genes OCT4, REX1, and NANOG; immunofluorescence-positive staining was presented for OCT4 and NANOG (all groups), SOX2 (groups 5% O-2 and 20% to 5% O-2), and TRA-1-60, TRA-1-81, and SSEA-1 (only in 20% O-2); they formed embryoid bodies; and there is spontaneous differentiation in mesoderm, endoderm, and ectoderm embryonic germ layers. In addition to the differences in immunofluorescence analysis results, the eiPSC colonies generated at 20% O-2 presented a more compact morphology with a well-defined border than cells cultured in 5% O-2 and 20% to 5% O-2. Significant differences were also observed in the expression of genes related to glucose metabolism, mitochondrial fission, and hypoxia (GAPDH, GLUT3, MFN1, HIF1 alpha, and HIF2 alpha), after reprogramming. Our results show that the derivation of eiPSCs was not impaired by aging. Additionally, this study is the first to compare high and low oxygen cultures of eiPSCs, showing the generation of pluripotent cells with different profiles. Under the tested conditions, the lower oxygen tension did not favor the pluripotency of eiPSCs. This study shows that the impact of oxygen atmosphere has to be considered when culturing eiPSCs, as this condition influences the pluripotency characteristics. (AU)

FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
Grantee:Dimas Tadeu Covas
Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
FAPESP's process: 18/04009-6 - Effects of low oxygen tension in the reprogramming efficiency and manutention of induced pluripotent stem cells in equine
Grantee:Raquel Vasconcelos Guimarães de Castro
Support Opportunities: Scholarships in Brazil - Doctorate
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 Opportunities: Research Grants - Young Investigators Grants