Human Induced Pluripotent Stem Cell Production and Expansion from Blood using a Non‐Integrating Viral Reprogramming Vector. Issue 1 (16th April 2018)

• Record Type: Journal Article
• Authors: Sharma, Arun; Mücke, Michael; Seidman, Christine E.
• Editors: Ausubel, Frederick M.; Brent, Roger; Kingston, Robert E.; Moore, David D.; Seidman, J.G.; Smith, John A.; Struhl, Kevin
• Is Part Of: Current protocols in molecular biology. Volume 121:Issue 1(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-04-16
• Subjects: cell fate -- iPSCs -- PBMC -- pluripotent stem cells -- reprogramming -- Sendai virus; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/cpmb.58
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: 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.