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

Sharma Arun | Zendy; Mücke Michael | Zendy; Seidman Christine E. | Zendy

Open Access

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

Author(s) -

Sharma Arun,

Mücke Michael,

Seidman Christine E.

Publication year - 2018

Publication title -

current protocols in molecular biology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.533

H-Index - 42

eISSN - 1934-3647

pISSN - 1934-3639

DOI - 10.1002/cpmb.58

Subject(s) - reprogramming , induced pluripotent stem cell , peripheral blood mononuclear cell , sendai virus , biology , microbiology and biotechnology , viral vector , virology , computational biology , immunology , cell , virus , embryonic stem cell , genetics , in vitro , recombinant dna , gene

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.

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