In vitro induced pluripotency from urine-derived cells in porcine.

Methods: The UDCs were reprogrammed in vitro using human or murine octamer-binding transcription factor 4 (OCT4), SRY-box2 (SOX2), Kruppel-like factor 4 (KLF4), and C-MYC, and cultured with basic fibroblast growth factor (bFGF) supplementation. To characterize the putative porcine iPSCs three clonal lineages were submitted to immunocytochemistry for alkaline phosphatase (AP), OCT4, SOX2, NANOG, TRA1 81 and SSEA 1 detection. Endogenous transcripts related to the pluripotency (OCT4, SOX2 and NANOG) were analyzed via reverse transcription quantitative real-time polymerase chain reaction in different time points during the culture, and all three lineages formed embryoid bodies (EBs) when cultured in suspension without bFGF supplementation. Results: The UDCs were isolated from swine urine samples and when at passage 2 submitted to in vitro reprogramming. Colonies of putative iPSCs were obtained only from UDCs transduced with the murine factors (mOSKM), but not from human factors (hOSKM). Three clonal lineages were isolated and further cultured for at least 28 passages, all the lineages were positive for AP detection, the OCT4, SOX2, NANOG markers, albeit the immunocytochemical analysis also revealed heterogeneous phenotypic profiles among lineages and passages for NANOG and SSEA1, similar results were observed in the abundance of the endogenous transcripts related to pluripotent state. All the clonal lineages when cultured in suspension without bFGF were able to form EBs expressing ectoderm and mesoderm layers transcripts. Conclusion: For the first time UDCs were isolated in the swine model and reprogrammed into a pluripotent-like state, enabling new numerous applications in both human or veterinary regenerative medicine.

Bibliographic Details

Main Authors:	RECCHIA, K., MACHADO, L. S., BOTIGELLI, R. C., PIERI, N. C. G., BARBOSA, G., CASTRO, R. V. G. de, MARQUES, M. G., PESSÔA, L. V. de F., FANTINATO NETO, P., MEIRELLES, F. V., SOUZA, A. F. de, MARTINS, S. M. M. K., BRESSAN, F. F.
Other Authors:	KAIANA RECCHIA, Universidade de São Paulo; LUCAS SIMÕES MACHADO, Universidade de São Paulo; RAMON CESAR BOTIGELLI, Universidade Estadual Paulista; NAIRA CAROLINE GODOY PIERI, Universidade de São Paulo; GABRIELA BARBOSA, Universidade de São Paulo; RAQUEL VASCONCELOS GUIMARÃES DE CASTRO, Universidade de São Paulo; MARIANA GROKE MARQUES, CNPSA; LAÍS VICARI DE FIGUEIREDO PESSÔA, Universidade de São Paulo; PAULO FANTINATO NETO, Universidade de São Paulo; FLÁVIO VIEIRA MEIRELLES, Universidade de São Paulo; ALINE FERNANDA DE SOUZA, Universidade de São Paulo; SIMONE MARIA MASSAMI KITAMURA MARTINS, Universidade de São Paulo; FABIANA FERNANDES BRESSAN, Universidade de São Paulo.
Format:	Artigo de periódico biblioteca
Language:	Ingles English
Published:	World Journal of Stem Cells, v. 14, n. 3, p. 231-244, 2022. 2022-12-21
Subjects:	Células-tronco, Pluripotência, IPSC, Noninvasive, Pluripotency, Reprogramming, Suíno, Urina, Induced pluripotent stem cells, Urine, Swine,
Online Access:	http://www.alice.cnptia.embrapa.br/alice/handle/doc/1150134 https://doi.org/10.4252/wjsc.v14.i3.231
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