Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression.
Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression.
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DOI：
10.1038/nprot.2012.121
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发表时间：
2012-11
期刊：
影响因子：
14.8
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中科院分区：
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几种人类出生后的体细胞类型已经成功地被重新编程为诱导多能干细胞(IPSCs)。与其他类型的细胞相比，血液单个核细胞(MNC)具有许多优势。它们很容易从脐带血(CB)或成人外周血(PB)中分离出来，可以新鲜使用或冷冻后使用。较短的培养时间可以实现更有效的重编程，血液MNC在14天内形成IPSC克隆，而年龄匹配的成纤维细胞在28天内形成IPSC克隆。短暂培养的血液单个核细胞的优势可能是由于良好的表观遗传学特征和基因表达模式。来自成人的血细胞，特别是经常从间歇性激活的血液干细胞中补充的非淋巴样细胞，在体内寿命很短，可能比皮肤成纤维细胞含有的体细胞突变更少，后者随着时间的推移更容易受到环境诱变剂的影响。在这里，我们描述了一种详细的、有效的、有效地从血液中的MNC中通过质粒载体产生无整合的人IPSCs的方案。
Several human postnatal somatic cell types have been successfully reprogrammed to induced pluripotent stem cells (iPSCs). Blood mononuclear cells (MNCs) offer several advantages compared with other cell types. They are easily isolated from umbilical cord blood (CB) or adult peripheral blood (PB), and can be used fresh or after freezing. A short culture allows for more efficient reprogramming, with iPSC colonies forming from blood MNCs in 14 d, compared with 28 d for age-matched fibroblastic cells. The advantages of briefly cultured blood MNCs may be due to favorable epigenetic profiles and gene expression patterns. Blood cells from adults, especially nonlymphoid cells that are replenished frequently from intermittently activated blood stem cells, are short-lived in vivo and may contain less somatic mutations than skin fibroblasts, which are more exposed to environmental mutagens over time. We describe here a detailed, validated protocol for effective generation of integration-free human iPSCs from blood MNCs by plasmid vectors.