Machado Lucas Simões, Borges Camila Martins, de Lima Marina Amaro, Sangalli Juliano Rodrigues, Therrien Jacinthe, Pessôa Laís Vicari de Figueiredo, Fantinato Neto Paulo, Perecin Felipe, Smith Lawrence Charles, Meirelles Flavio Vieira, Bressan Fabiana Fernandes
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.
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-900, SP, Brazil.
Biomedicines. 2023 Sep 19;11(9):2577. doi: 10.3390/biomedicines11092577.
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 or , or both, were analyzed regarding the expression of pluripotency factors and imprinted genes and , and used for in vitro reprogramming. The expression of the 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.
多能性相关转录因子或核移植介导的细胞重编程机制似乎由相似的途径介导,研究OCT4和SOX2在这两个过程中的作用可能有助于阐明多能性的相关机制。对表达外源性OCT4、SOX2或两者的牛成纤维细胞进行多能性因子和印记基因XIST和PEG10表达分析,并用于体外重编程。在对照分选组中,XIST基因的表达增加,当细胞未进行细胞分选时可获得假定的类诱导多能干细胞。当将表达OCT4、SOX2或不表达(对照)的分选细胞用作体细胞核移植的供体细胞时,SOX2组与对照组、OCT4组与对照组的融合率分别为60.0%对64.95%和70.53%对67.24%;裂解率分别为66.66%对81.68%和86.47%对85.18%;囊胚率分别为33.05%对44.15%和52.06%对44.78%。这些结果表明,与对照细胞相比,核移植产生的胚胎体外发育能力相似,无论供体细胞呈现出何种多能性相关基因表达谱;然而,细胞分选后诱导重编程受到损害。