Ankara University, Stem Cell Institute, Ankara, Turkey.
Ankara University, Faculty of Medicine, Department of Biophysics, Ankara, Turkey.
Methods Mol Biol. 2024;2835:99-110. doi: 10.1007/978-1-0716-3995-5_9.
Induced pluripotent stem cells (iPSCs) are generated through the reprogramming of somatic cells to an embryonic-like state by activating specific genes. They closely resemble embryonic stem cells (ESCs), in various aspects, including the expression of key stem cell genes, potency, and differentiation capabilities. iPSCs can be derived from various cell types such as fibroblasts, keratinocytes, and peripheral blood mononuclear cells (PBMCs). The ease of obtaining origin cells through non-invasive methods simplifies the generation of human iPSCs. Therefore, PBMCs are commonly preferred, with erythroid progenitor cells (EPCs) obtained through EPC enrichment being used as origin cells in this protocol. The EPC enrichment performed in this protocol not only reduces costs but also increases efficiency by enhancing the percentage of reprogrammable cells with progenitor characteristics. Human iPSCs are incredibly valuable for in vitro research, cell therapy, drug discovery, and tissue engineering. The outlined procedures below provide a general framework for inducing iPSCs from erythroid progenitor cells, pluripotency confirmation experiments, and cultivating them for downstream experiments.
诱导多能干细胞(iPSCs)是通过激活特定基因将体细胞重编程为类似胚胎的状态而产生的。它们在许多方面与胚胎干细胞(ESCs)相似,包括关键干细胞基因的表达、多能性和分化能力。iPSCs 可以源自多种细胞类型,如成纤维细胞、角质细胞和外周血单核细胞(PBMCs)。通过非侵入性方法获得起始细胞的简便性简化了人 iPSCs 的生成。因此,通常首选 PBMCs,并且在此方案中使用通过 EPC 富集获得的红系祖细胞(EPC)作为起始细胞。该方案中进行的 EPC 富集不仅通过增强具有祖细胞特征的可重编程细胞的百分比来降低成本,而且还提高了效率。人 iPSCs 对于体外研究、细胞治疗、药物发现和组织工程非常有价值。下面概述的程序提供了从红系祖细胞诱导 iPSCs、多能性确认实验以及为下游实验培养它们的一般框架。