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©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
In vitro induced pluripotency from urine-derived cells in porcine
Kaiana Recchia, Lucas Simões Machado, Ramon Cesar Botigelli, Naira Caroline Godoy Pieri, Gabriela Barbosa, Raquel Vasconcelos Guimarães de Castro, Mariana Groke Marques, Laís Vicari de Figueiredo Pessôa, Paulo Fantinato Neto, Flávio Vieira Meirelles, Aline Fernanda de Souza, Simone Maria Massami Kitamura Martins, Fabiana Fernandes Bressan
Kaiana Recchia, Lucas Simões Machado, Flávio Vieira Meirelles, Fabiana Fernandes Bressan, Department of Surgery, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, Pirassununga 13635-000, São Paulo, Brazil
Ramon Cesar Botigelli, Department of Pharmacology and Biotechnology, Institute of Bioscience, São Paulo State University, Botucatu 18618-689, São Paulo, Brazil
Naira Caroline Godoy Pieri, Gabriela Barbosa, Raquel Vasconcelos Guimarães de Castro, Laís Vicari de Figueiredo Pessôa, Paulo Fantinato Neto, Flávio Vieira Meirelles, Aline Fernanda de Souza, Fabiana Fernandes Bressan, Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-000, São Paulo, Brazil
Mariana Groke Marques, Embrapa Suínos e Aves, Empresa Brasileira de Pesquisa Agropecuária, Concordia 89715-899, Santa Catarina, Brazil
Simone Maria Massami Kitamura Martins, Department of Animal Sciences, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-000, São Paulo, Brazil
Author contributions: Recchia K and Bressan FF conceptualized the experiments and wrote the manuscript; Recchia K, Machado LS, Barbosa G, Marques MG and Martins SMMK collected and processed samples; Recchia K, Botigelli RC, Pieri NCG, de Castro RVG, Meirelles FV, de Souza AF, Pessôa LVF, Fantinato Neto P and Bressan FF collected data and interpreted the collected data; all authors revised the manuscript.
Supported by the São Paulo Research Foundation (FAPESP) - Brazil (financial support grants #2015/26818-5, #2013/08135-2, #2019/02811-2); Coordination of Superior Level Staff Improvement (CAPES 23038.006964/2014-43 and financial code 001) and National Council for Scientific and Technological Development (CNPq 433133/2018-0).
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo.
Institutional animal care and use committee statement: All procedures were performed following the National Council for Control of Animal Experimentation (CONCEA) rules and were approved by the Ethics Committee on Animal Experimentation of the Faculty of Animal Science and Food Engineering and Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo (protocols 6372070119 and 7051150717).
Conflict-of-interest statement: The authors declare that there are no conflicts of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Fabiana Fernandes Bressan, DVM, Professor, Department of Surgery, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, Av Duque de Caxias Norte 225, Pirassununga 13635-000, São Paulo, Brazil.
fabianabressan@usp.br
Received: August 23, 2021
Peer-review started: August 23, 2021
First decision: October 3, 2021
Revised: October 11, 2021
Accepted: February 15, 2022
Article in press: February 15, 2022
Published online: March 26, 2022
Processing time: 213 Days and 19.5 Hours
ARTICLE HIGHLIGHTS
Research background
Induced pluripotent stem cells (iPSCs) derived from large animal models can greatly contribute to translational medicine and also to animal production, although robust and safe protocols are still uncommon. Cellular reprogramming of urine derived cells presents great advantages for iPSCs use in regenerative medicine due to the easy collection, injury and stress free, and is herein described for the first time in large animals.
Research motivation
The porcine iPSCs generation is promising for both translational medicine and animal production; and iPSCs derived from a noninvasive cell source would greatly contribute to its large-scale use, especially for in vivo autologous purposes using large animal models.
Research objectives
Isolate cells from porcine urine and generate iPSCs through their transduction with Yamanaka’s human or murine factors.
Research methods
We isolated urine-derived cells (UDCs), which were reprogrammed in vitro into pluripotent cells. The porcine induced pluripotent cells generated were investigated regarding morphology, markers and endogenous transcripts related to the pluripotency.
Research results
From the porcine urine samples we isolated the UDCs, and colonies were formed when murine factors were used in the reprogramming. Endogenous pluripotent markers were detected in all three isolated lineages, in different time points during in vitro culture, and were able to differentiate into embryoid bodies (EBs) with mesoderm and ectoderm transcripts.
Research conclusions
In an unprecedented way, UDCs were isolated from noninvasive collection and reprogrammed into a pluripotent state using murine factors, the cells formed colonies presenting the expected characteristics, such as colonies with limited borders, transcripts and markers related to the pluripotency, and ability to differentiate into EBs.
Research perspectives
As we reported here, iPSCs can be derived from an easy collection and noninvasive source in the porcine model, and with our methodology represents a novel tool for iPSCs production in large animals and biomedical models of regenerative or translational medicine.
