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从外周血内皮祖细胞中诱导多能干细胞的无饲养层培养法

Feeder-independent derivation of induced-pluripotent stem cells from peripheral blood endothelial progenitor cells.

作者信息

Chang Wing Y, Lavoie Jessie R, Kwon Sarah Y, Chen Zhaoyi, Manias Janet L, Behbahani John, Ling Vicki, Kandel Rita A, Stewart Duncan J, Stanford William L

机构信息

Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, Canada.

出版信息

Stem Cell Res. 2013 Mar;10(2):195-202. doi: 10.1016/j.scr.2012.11.006. Epub 2012 Dec 3.

Abstract

Induced-pluripotent stem cells (iPSCs) are a potential alternative cell source in regenerative medicine, which includes the use of differentiated iPSCs for cell therapies to treat coronary artery and/or peripheral arterial diseases. Late-outgrowth endothelial progenitor cells (late-EPCs) are a unique primary cell present in peripheral blood that exhibit high proliferative capacity, are being used in a wide variety of clinical trials, and have the ability to differentiate into mature endothelial cells. The objective of this study was to reprogram peripheral blood-derived late-EPCs to a pluripotent state under feeder-free and defined culture conditions. Late-EPCs that were retrovirally transduced with OCT4, SOX2, KLF4, c-MYC, and iPSC colonies were derived in feeder-free and defined media conditions. EPC-iPSCs expressed pluripotent markers, were capable of differentiating to cells from all three germ-layers, and retained a normal karyotype. Transcriptome analyses demonstrated that EPC-iPSCs exhibit a global gene expression profile similar to human embryonic stem cells (hESCs). We have generated iPSCs from late-EPCs under feeder-free conditions. Thus, peripheral blood-derived late-outgrowth EPCs represent an alternative cell source for generating iPSCs.

摘要

诱导多能干细胞(iPSC)是再生医学中一种潜在的替代细胞来源,其中包括使用分化的iPSC进行细胞治疗,以治疗冠状动脉和/或外周动脉疾病。晚期内皮祖细胞(late-EPC)是外周血中存在的一种独特的原代细胞,具有高增殖能力,正在广泛用于各种临床试验,并且能够分化为成熟的内皮细胞。本研究的目的是在无饲养层和明确的培养条件下,将外周血来源的晚期内皮祖细胞重编程为多能状态。用OCT4、SOX2、KLF4、c-MYC进行逆转录病毒转导的晚期内皮祖细胞和iPSC集落是在无饲养层和明确的培养基条件下获得的。EPC-iPSC表达多能性标志物,能够分化为来自所有三个胚层的细胞,并保持正常的核型。转录组分析表明,EPC-iPSC表现出与人类胚胎干细胞(hESC)相似的整体基因表达谱。我们在无饲养层条件下从晚期内皮祖细胞生成了iPSC。因此,外周血来源的晚期内皮祖细胞代表了一种生成iPSC的替代细胞来源。

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