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.
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References
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Acknowledgements
The authors thank Dr. Thomas Moreau for expert advice and support. The authors also acknowledge the contribution of “Fundação para a Ciência e a Tecnologia” (SFRH/BD/69033/2010) and Cambridge Hospitals National Institute for Health Research Biomedical Research Center and ERC starting grant Relieve-IMDs.
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© 2015 Springer Science+Business Media New York
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Soares, F.A.C., Pedersen, R.A., Vallier, L. (2015). Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus. In: Turksen, K., Nagy, A. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 1357. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_202
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DOI: https://doi.org/10.1007/7651_2015_202
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3054-8
Online ISBN: 978-1-4939-3055-5
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