Title:
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Simple generation of human induced pluripotent stem cells using poly-β-amino esters as the non-viral gene delivery system
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Author:
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Borrós i Gómez, Salvador; Ramos Pérez, Víctor; Gutiérrez, Jordán; Izpisúa Belmonte, J. C.; Montserrat, Núria; Garreta, Elena; González, Federico; Eguizábal, Cristina
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Other authors:
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Universitat Ramon Llull. IQS |
Abstract:
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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. |
Publication date:
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2011-04 |
Subject (UDC):
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576 - Biologia cel·lular i subcel·lular. Citologia |
Subject(s):
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Genètica humana Poliamines Proteïnes--Síntesi Cèl·lules mare Nanopartícules Esters Transfecció Human genetics Polyaminers Protein synthesis Stem cells iPS cells Reprogramming Transfection |
Rights:
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© The American Society for Biochemistry and Molecular Biology. Tots els drets reservats |
Pages:
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13 p. |
Document type:
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Article Article - Published version |
DOI:
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https://doi.org/10.1074/jbc.m110.168013
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Published by:
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The American Society for Biochemistry and Molecular Biology
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Publish at:
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Journal of Biological Chemistry. Vol.286,n.14 (2011),p.12417-12428
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