Recchia Kaiana, Pessôa Laís Vicari de Figueiredo, Pieri Naira Caroline Godoy, Pires Pedro Ratto Lisboa, Bressan Fabiana Fernandes
Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, São Paulo 05508-270, SP, Brazil.
Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Av. Duque de Caxias Norte, 225, Pirassununga 13635-900, SP, Brazil.
Life (Basel). 2022 Jul 28;12(8):1139. doi: 10.3390/life12081139.
Induced pluripotent stem cells (iPSCs) have been considered an essential tool in stem cell research due to their potential to develop new therapies and technologies and answer essential questions about mammalian early development. An important step in generating iPSCs is selecting their precursor cell type, influencing the reprogramming efficiency and maintenance in culture. In this study, we aim to characterize bovine mesenchymal cells from adipose tissue (bAdMSCs) and fetal fibroblasts (bFFs) and to compare the reprogramming efficiency of these cells when induced to pluripotency. The cells were characterized by immunostaining (CD90, SSEA1, SSEA3, and SSEA4), induced differentiation in vitro, proliferation rates, and were subjected to cell reprogramming using the murine OSKM transcription factors. The bFFs presented morphological changes resembling pluripotent cells after reprogramming and culture with different supplementation, and putative iPSCs were characterized by immunostaining (OCT4, SOX2, NANOG, and AP). In the present study, we demonstrated that cell line origin and cellular proliferation rate are determining factors for reprogramming cells into pluripotency. The generation of biPSCs is a valuable tool to improve both translational medicine and animal production and to study the different supplements required to maintain the pluripotency of bovine cells in vitro.
诱导多能干细胞(iPSC)因其具有开发新疗法和技术的潜力以及解答有关哺乳动物早期发育的关键问题,而被视为干细胞研究中的一项重要工具。生成iPSC的一个重要步骤是选择其前体细胞类型,这会影响重编程效率以及在培养中的维持。在本研究中,我们旨在对来自脂肪组织的牛间充质细胞(bAdMSC)和胎儿成纤维细胞(bFF)进行表征,并比较这些细胞被诱导为多能性时的重编程效率。通过免疫染色(CD90、SSEA1、SSEA3和SSEA4)、体外诱导分化、增殖率对细胞进行表征,并使用小鼠OSKM转录因子对细胞进行重编程。重编程并在不同补充剂条件下培养后,bFF呈现出类似于多能细胞的形态变化,通过免疫染色(OCT4、SOX2、NANOG和AP)对假定的iPSC进行表征。在本研究中,我们证明细胞系来源和细胞增殖率是将细胞重编程为多能性的决定因素。生物iPSC的生成是改善转化医学和动物生产以及研究体外维持牛细胞多能性所需的不同补充剂的宝贵工具。