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Published in:

2015 | Online First | Chapter

Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus

Authors : Filipa A. C. Soares, Roger A. Pedersen, Ludovic Vallier

Published in: Methods in Molecular Biology™

Publisher: Springer New York

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Abstract

This protocol describes the efficient isolation of peripheral blood mononuclear cells from circulating blood via density gradient centrifugation and subsequent generation of integration-free human induced pluripotent stem cells. Peripheral blood mononuclear cells are cultured for 9 days to allow expansion of the erythroblast population. The erythroblasts are then used to derive human induced pluripotent stem cells using Sendai viral vectors, each expressing one of the four reprogramming factors Oct4, Sox2, Klf4, and c-Myc.
Literature
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Yang W, Mills JA, Sullivan S, Liu Y, French DL, Gadue P (2012) iPSC Reprogramming from human peripheral blood using sendai virus mediated gene transfer. In: StemBook (ed) The Stem Cell Research Community, StemBook
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Chou B-K, Mali P, Huang X, Ye Z, Dowey SN, Resar LMS, Zou C, Zhang YA, Tong J, Cheng L (2011) Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res 21(3):518–529PubMedPubMedCentralCrossRef
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van den Akker E, Satchwell TJ, Pellegrin S, Daniels G, Toye AM (2010) The majority of the in vitro erythroid expansion potential resides in CD34(-)cells, outweighing the contribution of CD34(+) cells and significantly increasing the erythroblast yield from peripheral blood samples. Haematologica 95(9):1594–1598PubMedPubMedCentralCrossRef
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Leberbauer C, Boulmé F, Unfried G, Huber J, Beug H, Müllner EW (2005) Different steroids co-regulate long-term expansion versus terminal differentiation in primary human erythroid progenitors. Blood 105(1):85–94PubMedCrossRef
Metadata
Title
Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus
Authors
Filipa A. C. Soares
Roger A. Pedersen
Ludovic Vallier
Copyright Year
2015
Publisher
Springer New York
DOI
https://doi.org/10.1007/7651_2015_202