Generation of adult human induced pluripotent stem cells using nonviral minicircle DNA vectors

被引：142

作者：

Narsinh, Kazim H. ^{[1
,2
,3
]}

Jia, Fangjun ^{[1
,2
]}

Robbins, Robert C. ^{[4
]}

Kay, Mark A. ^{[5
,6
]}

Longaker, Michael T. ^{[4
,7
]}

Wu, Joseph C. ^{[1
,2
,7
]}

机构：

[1] Stanford Univ, Dept Med, Sch Med, Stanford, CA 94305 USA

[2] Stanford Univ, Dept Radiol, Sch Med, Stanford, CA 94305 USA

[3] Univ Calif San Diego, Sch Med, La Jolla, CA 92093 USA

[4] Stanford Univ, Dept Surg, Sch Med, Stanford, CA 94305 USA

[5] Stanford Univ, Dept Pediat, Sch Med, Stanford, CA 94305 USA

[6] Stanford Univ, Dept Genet, Sch Med, Stanford, CA 94305 USA

[7] Stanford Univ, Inst Stem Cell Biol & Regenerat Med, Sch Med, Stanford, CA 94305 USA

来源：

NATURE PROTOCOLS | 2011年 / 6卷 / 01期

基金：

美国国家卫生研究院;

关键词：

INDUCTION; FIBROBLASTS; PERSISTENT;

D O I：

10.1038/nprot.2010.173

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Human induced pluripotent stem cells (hiPSCs) derived from patient samples have tremendous potential for innovative approaches to disease pathology investigation and regenerative medicine therapies. However, most hiPSC derivation techniques use integrating viruses, which may leave residual transgene sequences as part of the host genome, thereby unpredictably altering cell phenotype in downstream applications. In this study, we describe a protocol for hiPSC derivation by transfection of a simple, nonviral minicircle DNA construct into human adipose stromal cells (hASCs). Minicircle DNA vectors are free of bacterial DNA and thus capable of high expression in mammalian cells. Their repeated transfection into hASCs, abundant somatic cell sources that are amenable to efficient reprogramming, results in transgene-free hiPSCs. This protocol requires only readily available molecular biology reagents and expertise, and produces hiPSC colonies from an adipose tissue sample in similar to 4 weeks.

引用

页码：78 / 88

页数：11

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