Exogenous OCT4 and SOX2 Contribution to In Vitro Reprogramming in Cattle
Lucas Simões Machado,
Camila Martins Borges,
Marina Amaro de Lima,
Juliano Rodrigues Sangalli,
Jacinthe Therrien,
Laís Vicari de Figueiredo Pessôa,
Paulo Fantinato Neto,
Felipe Perecin,
Lawrence Charles Smith,
Flavio Vieira Meirelles,
Fabiana Fernandes Bressan
Affiliations
Lucas Simões Machado
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Camila Martins Borges
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Marina Amaro de Lima
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Juliano Rodrigues Sangalli
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-900, SP, Brazil
Jacinthe Therrien
Centre de Recherche en Reproduction et Fertilité, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC J2S 7C6, Canada
Laís Vicari de Figueiredo Pessôa
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-900, SP, Brazil
Paulo Fantinato Neto
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-900, SP, Brazil
Felipe Perecin
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-900, SP, Brazil
Lawrence Charles Smith
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Flavio Vieira Meirelles
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Fabiana Fernandes Bressan
Post-Graduate Program of Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo 05508-270, SP, Brazil
Mechanisms of cell reprogramming by pluripotency-related transcription factors or nuclear transfer seem to be mediated by similar pathways, and the study of the contribution of OCT4 and SOX2 in both processes may help elucidate the mechanisms responsible for pluripotency. Bovine fibroblasts expressing exogenous OCT4 or SOX2, or both, were analyzed regarding the expression of pluripotency factors and imprinted genes H19 and IGF2R, and used for in vitro reprogramming. The expression of the H19 gene was increased in the control sorted group, and putative iPSC-like cells were obtained when cells were not submitted to cell sorting. When sorted cells expressing OCT4, SOX2, or none (control) were used as donor cells for somatic cell nuclear transfer, fusion rates were 60.0% vs. 64.95% and 70.53% vs. 67.24% for SOX2 vs. control and OCT4 vs. control groups, respectively; cleavage rates were 66.66% vs. 81.68% and 86.47% vs. 85.18%, respectively; blastocyst rates were 33.05% vs. 44.15% and 52.06% vs. 44.78%, respectively. These results show that the production of embryos by NT resulted in similar rates of in vitro developmental competence compared to control cells regardless of different profiles of pluripotency-related gene expression presented by donor cells; however, induced reprogramming was compromised after cell sorting.
