Division of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, MD, USA.
Methods Mol Biol. 2022;2454:381-396. doi: 10.1007/7651_2021_372.
Induced pluripotent stem cells (iPSCs) offer the potential to generate tissue cells with donor diversity therefore promising to have widespread applications in regenerative medicine, disease modeling, drug discovery, and toxicity testing. Several somatic cell types have been utilized, with varying efficiencies, as source cells for the reprogramming of iPSCs. Recently, it has been reported that endothelial progenitor cells (EPCs) derived from umbilical cord blood (CB) or adult peripheral blood (PB) afford a practical and efficient cellular substrate for iPSC generation, and possess several advantages over other cell types. In this chapter, we describe a protocol that covers all steps of reprogramming iPSCs from blood-derived EPCs, including (1) isolation of mononuclear cells (MNCs) from blood samples, (2) derivation of EPCs from MNCs, and (3) generation of iPSCs from EPCs. The final step of reprogramming EPCs into iPSCs is achieved through ectopic expression of four transcription factors, OCT4, KLF4, SOX2, and c-MYC, using self-replicative RNA (srRNA) technology.
诱导多能干细胞 (iPSC) 具有产生具有供体多样性的组织细胞的潜力，因此有望在再生医学、疾病建模、药物发现和毒性测试中得到广泛应用。已经利用了几种体细胞类型作为重编程 iPSC 的源细胞，其效率各不相同。最近有报道称，源自脐带血 (CB) 或成人外周血 (PB) 的内皮祖细胞 (EPC) 为 iPSC 的产生提供了一种实用且高效的细胞基质，并且与其他细胞类型相比具有几个优势。在本章中，我们描述了一种从血液衍生的 EPC 中重编程 iPSC 的方案，包括：（1）从血液样本中分离单核细胞 (MNC)，（2）从 MNC 中衍生 EPC，以及（3）从 EPC 中生成 iPSC。通过使用自我复制 RNA (srRNA) 技术过表达四个转录因子 OCT4、KLF4、SOX2 和 c-MYC，EPC 被重编程为 iPSC。
