Human Induced Pluripotent Stem Cell Production and Expansion from Blood using a Non-Integrating Viral Reprogramming Vector

We describe a method to transform blood lymphocytes into human-induced pluripotent stem cells by delivering four transcription factors with a non-integrative virus. Using human peripheral blood mononuclear cells (PBMCs) as the source cell type for hiPSC reprogramming is advantageous since blood samp...

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Bibliographic Details
Published inCurrent protocols in molecular biology (Print) Vol. 122; no. 1; p. e58
Main Authors Sharma, Arun, Mücke, Michael, Seidman, Christine E
Format Journal Article
LanguageEnglish
Published United States 01.04.2018
Subjects
Blood Cells - metabolism
Cell Proliferation
Cell Separation
Genetic Vectors - metabolism
Humans
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - metabolism
Sendai virus - genetics
Virus Integration
reprogramming
iPSCs
pluripotent stem cells
Sendai virus
PBMC
cell fate
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Summary:We describe a method to transform blood lymphocytes into human-induced pluripotent stem cells by delivering four transcription factors with a non-integrative virus. Using human peripheral blood mononuclear cells (PBMCs) as the source cell type for hiPSC reprogramming is advantageous since blood samples are rapidly and safely obtained from nearly-all subjects. Reprogramming factors needed to make hiPSCs are introduced by infecting the PBMCs with non-integrating Sendai virus vectors. Reprogrammed cells can subsequently be quickly expanded for downstream use. In this unit, we present current protocols for the isolation of PBMCs from a small sample of human blood and subsequent viral reprogramming and expansion of PBMCs into hiPSCs. © 2018 by John Wiley & Sons, Inc.
ISSN:1934-3647
DOI:10.1002/cpmb.58