Generation of induced pluripotent stem cells from a small amount of human peripheral blood using a combination of activated T cells and Sendai virus

Department of Cardiology, Keio University School of Medicine, Tokyo, Japan.
Nature Protocol (Impact Factor: 8.36). 03/2012; 7(4):718-28. DOI: 10.1038/nprot.2012.015
Source: PubMed

ABSTRACT Induced pluripotent stem cells (iPSCs) have become important cell sources for genetic disease models, and they have the potential to be cell sources for future clinical therapies. However, invasive tissue sampling reduces the number of candidates who consent to donate cells for iPSC generation. In addition, integrated transgenes can potentially insert at inappropriate points in the genome, and in turn have a direct oncogenic effect. Technical modifications using a combination of activated T cells and a temperature-sensitive mutant of Sendai virus (SeV) can avoid invasive tissue sampling and residual transgene issues in generating iPSCs. Such advances may increase the number of consenting patients for cell donations. Here we present a detailed protocol for the generation of iPSCs from a small amount of human peripheral blood using a combination of activated T cells and mutant SeV encoding human OCT3/4, SOX2, KLF4 and c-MYC; T cell-derived iPSCs can be generated within 1 month of blood sampling.

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    • "Most of the commonly-used source tissues or cells for iPSC production are skin biopsies and blood cells, and there are multiple ways of delivering the pluripotent re-programming factors (Oct3/4, Klf4, Sox2, and c-Myc), which have been reviewed extensively elsewhere [94] . The optimal way to generate iPSC lines would be free of virus integration, e.g., by Sendai virus [95] , that has been used by the NIMH stem cell center ( to establish a resource of iPSC lines for psychiatric research. "
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