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

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2021 Jul 2

PMID 34204517

Citations 9

Authors

Brendon Willian Bessi

Ramon Cesar Botigelli

Naira Caroline Godoy Pieri

Lucas Simoes Machado

Jessica Brunhara Cruz

Pamela de Moraes

Aline Fernanda de Souza

Kaiana Recchia

Gabriela Barbosa

Raquel Vasconcelos Guimaraes de Castro

Marcelo Fabio Gouveia Nogueira

Fabiana Fernandes Bressan

Affiliations

Soon will be listed here.

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 , and . However, expression was decreased throughout the passages without the influence of oxygen tension. and 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.

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