Simple generation of human induced pluripotent stem cells using poly-β-amino esters as the non-viral gene delivery system
Author
Other authors
Publication date
2011-04Abstract
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.
Document Type
Article
Published version
Language
English
Subject (CDU)
576 - Cellular and subcellular biology. Cytology
Keywords
Pages
13 p.
Publisher
The American Society for Biochemistry and Molecular Biology
Is part of
Journal of Biological Chemistry. Vol.286,n.14 (2011),p.12417-12428
This item appears in the following Collection(s)
Rights
© The American Society for Biochemistry and Molecular Biology. Tots els drets reservats