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

• Published in: Cells (Basel, Switzerland) Vol. 10; no. 6; p. 1531
• Main Authors: Bessi, Brendon Willian, Botigelli, Ramon Cesar, Pieri, Naira Caroline Godoy, Machado, Lucas Simões, Cruz, Jessica Brunhara, de Moraes, Pamela, de Souza, Aline Fernanda, Recchia, Kaiana, Barbosa, Gabriela, de Castro, Raquel Vasconcelos Guimarães, Nogueira, Marcelo Fábio Gouveia, Bressan, Fabiana Fernandes
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 17-06-2021; MDPI
• Subjects: acquisition of pluripotency; Alkaline phosphatase; Amino acids; bovine iPSCs; Cattle; Cell culture; Cell self-renewal; Ectoderm; Endoderm; Fibroblast growth factor 2; Fibroblasts; Genetic engineering; Hypoxia; Immunofluorescence; In vitro fertilization; Kinases; Livestock; Medical research; Mesoderm; Oct-4 protein; oxygen; Oxygen tension; Penicillin; Phosphatase; Pluripotency; pluripotent state; Stem cells; Supplements; St Louis Missouri; United States > US
• DOI: 10.3390/cells10061531
• EISSN: 2073-4409

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.