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Use of poly(β-amino esters) as a non-viral means to induce pluripotency



From the Journal of Biological Chemistry

Since the generation of the first induced pluripotent stem cells (iPSC) from somatic cells was reported in 2006, various alternative ways of achieving pluripotency have been attempted in order to improve the safety and efficiency of the reprogramming process and of the resultant iPSC. The production of the viral particles commonly used to express the reprogramming factors is a time and labour intensive process and the risk of insertional mutagenesis is of significant clinical concern. From the Center of Regenerative Medicine of Barcelona now comes a report by Montserrat et al.1 which describes that human fibroblasts can successfully be reprogrammed using poly(β-amino esters) as the transfection reagent, avoiding the use of viral vectors entirely. The authors report that using serial transfection with poly(β-amino esters), which are biodegradable polymers that are easy to synthesise and have low toxicity, they can successfully transfect human fibroblasts with a CAG driven vector expressing reprogramming factors (Oct4, Sox2, Klf4 and c-Myc tagged with a GFP reporter gene) as a single polycistronic plasmid, to generate iPSC in 20-28 days. Moreover, they do this with higher efficiency than commercially available transfection reagents, an important consideration given the usually low transfection efficiency observed in human cells. Although this method does not remove the need for transgenes, alternative methods to transfect cells to achieve induced pluripotency, coupled with new advances in transgene free methods (perhaps for example with the use of microRNAs), will avoid potential complications associated with viral integration into the genome or the use of potentially oncogenic transgenes.



1 Montserrat N et al. ‘Simple generation of human induced Pluripotent stem cells using Poly(β-Amino Esters) as non-viral gene delivery system’ J Biol Chem. 2011 Feb 1.