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J Biol Chem. 2011 Apr 8;286(14):12417-28. doi: 10.1074/jbc.M110.168013. Epub 2011 Feb 1.

Simple generation of human induced pluripotent stem cells using poly-beta-amino esters as the non-viral gene delivery system.

Author information

1
Center of Regenerative Medicine of Barcelona, Doctor Aiguader 88, p7, 08003 Barcelona, Spain.

Abstract

Reprogramming of somatic cells to induced pluripotent stem (iPS) cells can be achieved by the delivery of a combination of transcription factors, including Oct4, Sox2, Klf4, and c-Myc. Retroviral and lentiviral vectors are commonly used to express these four reprogramming factors separately and obtain reprogrammed iPS cells. Although efficient and reproducible, these approaches involve the time-consuming and labor-intensive production of retroviral or lentiviral particles together with a high risk of working with potentially harmful viruses overexpressing potent oncogenes, such as c-Myc. Here, we describe a simple method to produce bona fide iPS cells from human fibroblasts using poly-β-amino esters as the transfection reagent for the delivery of a single CAG-driven polycistronic plasmid expressing Oct4, Sox2, Klf4, c-Myc, and a GFP reporter gene (OSKMG). We demonstrate for the first time that poly-β-amino esters can be used to deliver a single polycistronic reprogramming vector into human fibroblasts, achieving significantly higher transfection efficiency than with conventional transfection reagents. After a protocol of serial transfections using poly-β-amino esters, we report a simple methodology to generate human iPS cells from human fibroblasts avoiding the use of viral vectors.

PMID:
21285354
PMCID:
PMC3069445
DOI:
10.1074/jbc.M110.168013
[Indexed for MEDLINE]
Free PMC Article

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