Cattle In Vitro Induced Pluripotent Stem Cells Generated and Maintained in 5 or 20% Oxygen and Different Supplementation

Author(s):Show less -

Bessi, Brendon Willian ^[1] ; Botigelli, Ramon Cesar ^{[1, 2]} ; Pieri, Naira Caroline Godoy ^{[3, 1]} ; Machado, Lucas Simoes ^[1] ; Cruz, Jessica Brunhara ^[1] ; de Moraes, Pamela ^[1] ; de Souza, Aline Fernanda ^[1] ; Recchia, Kaiana ^[1] ; Barbosa, Gabriela ^[1] ; de Castro, Raquel Vasconcelos Guimaraes ^{[1, 4]} ; Nogueira, Marcelo Fabio Gouveia ^[5] ; Bressan, Fabiana Fernandes ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Anim Sci & Food Engn, Dept Vet Med, BR-13635000 Pirassununga - Brazil

^[2] Sao Paulo State Univ UNESP, Inst Biosci IBB, Dept Pharmacol, BR-18618689 Botucatu, SP - Brazil

^[3] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Anim Reprod, BR-05508270 Sao Paulo - Brazil

^[4] Sao Paulo State Univ UNESP, Fac Agr & Vet Sci, Dept Pathol Reprod & One Hlth, BR-14884900 Botucatu, SP - Brazil

^[5] Sao Paulo State Univ UNESP, Sch Sci Humanities & Languages, Dept Biol Sci, Sao Paulo - Brazil

• Document type: Journal article
• Source: CELLS; v. 10, n. 6 JUN 2021.
• Web of Science Citations: 0

Abstract

The event of cellular reprogramming into pluripotency is influenced by several factors, such as in vitro culture conditions (e.g., culture medium and oxygen concentration). Herein, bovine iPSCs (biPSCs) were generated in different levels of oxygen tension (5% or 20% of oxygen) and supplementation (bFGF or bFGF + LIF + 2i-bFL2i) to evaluate the efficiency of pluripotency induction and maintenance in vitro. Initial reprogramming was observed in all groups and bFL2i supplementation initially resulted in a superior number of colonies. However, bFL2i supplementation in low oxygen led to a loss of self-renewal and pluripotency maintenance. All clonal lines were positive for alkaline phosphatase; they expressed endogenous pluripotency-related genes SOX2, OCT4 and STELLA. However, expression was decreased throughout the passages without the influence of oxygen tension. GLUT1 and GLUT3 were upregulated by low oxygen. The biPSCs were immunofluorescence-positive stained for OCT4 and SOX2 and they formed embryoid bodies which differentiated in ectoderm and mesoderm (all groups), as well as endoderm (one line from bFL2i in high oxygen). Our study is the first to compare high and low oxygen environments during and after induced reprogramming in cattle. In our conditions, a low oxygen environment did not favor the pluripotency maintenance of biPSCs. (AU)

• 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
• FAPESP's process: 19/10751-0 - Trophectoderm complementation assay for pluripotent cells: a proposal for assessing pluripotency in cattle
• Grantee: Ramon Cesar Botigelli
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 16/16841-2 - Trophectoderm complementation assay for IPS cells: a proposal for assessing pluripotency in cattle
• Grantee: Ramon Cesar Botigelli
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 12/50533-2 - GIFT: genomic improvement of fertilization traits in Danish and Brazilian cattle
• Grantee: Marcelo Fábio Gouveia Nogueira
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/24520-7 - Epigenetic modification mechanisms are activated by reactive oxygen (ro) in bovine induced pluripotent stem cells
• Grantee: Brendon Willian Bessi
• Support Opportunities: Scholarships in Brazil - Scientific Initiation