Department of Innovative Technologies in Medicine & Dentistry, "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini, 31, 66100 Chieti, Italy.
Department of Psychological Health and Territorial Sciences, "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini, 31, 66100 Chieti, Italy.
Int J Mol Sci. 2024 Aug 23;25(17):9169. doi: 10.3390/ijms25179169.
The high mortality in the global population due to chronic diseases highlights the urgency to identify effective alternative therapies. Regenerative medicine provides promising new approaches for this purpose, particularly in the use of induced pluripotent stem cells (iPSCs). The aim of the work is to establish a new pluripotency cell line obtained for the first time by reprogramming human gingival mesenchymal stem cells (hGMSCs) by a non-integrating method. The hGMSC-derived iPS line characterization is performed through morphological analysis with optical and electron scanning microscopy and through the pluripotency markers expression evaluation in cytofluorimetry, immunofluorescence, and RT-PCR. To confirm the pluripotency of new hGMSC-derived iPS, the formation of embryoid bodies (EBs), as an alternative to the teratoma formation test, is studied in morphological analysis and through three germ layers' markers' expression in immunofluorescence and RT-PCR. At the end, a comparative study between parental hGMSCs and derived iPS cells is performed also for the extracellular vesicles (EVs) and their miRNA content. The new hGMSC-derived iPS line demonstrated to be pluripotent in all aspects, thus representing an innovative dynamic platform for personalized tissue regeneration.
由于慢性疾病导致的全球人口高死亡率突显了确定有效替代疗法的紧迫性。再生医学为此提供了有前途的新方法,特别是在诱导多能干细胞(iPSC)的使用方面。这项工作的目的是建立一种新的多能性细胞系,该细胞系首次通过非整合方法对人牙龈间充质干细胞(hGMSC)进行重编程获得。通过光学和电子扫描显微镜进行形态分析,并通过流式细胞术、免疫荧光和 RT-PCR 评估多能性标志物的表达,对 hGMSC 来源的 iPS 细胞系进行表征。为了确认新的 hGMSC 来源的 iPS 的多能性,通过形态分析研究类胚体(EB)的形成,作为畸胎瘤形成试验的替代方法,并通过免疫荧光和 RT-PCR 研究三个胚层标志物的表达。最后,还对亲本 hGMSC 和衍生的 iPS 细胞进行了比较研究,包括细胞外囊泡(EVs)及其 miRNA 含量。新的 hGMSC 来源的 iPS 系在各个方面均表现出多能性,因此代表了个性化组织再生的创新动态平台。