Human induced pluripotent stem cell production and expansion from blood using a non-integrating viral reprogramming vector
Authors
- A. Sharma
- M. Mücke
- C.E. Seidman
Journal
- Current Protocols in Molecular Biology
Citation
- Curr Protoc Mol Biol 122 (1): e58
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.