Generation of transgene-free human induced pluripotent stem cells from erythroblasts in feeder-free conditions

Sylvain Perriot; Mathieu Canales; Amandine Mathias; Renaud Du Pasquier

doi:10.1016/j.xpro.2022.101620

Generation of transgene-free human induced pluripotent stem cells from erythroblasts in feeder-free conditions

STAR Protoc. 2022 Aug 17;3(3):101620. doi: 10.1016/j.xpro.2022.101620. eCollection 2022 Sep 16.

Authors

Sylvain Perriot¹, Mathieu Canales², Amandine Mathias², Renaud Du Pasquier³

Affiliations

¹ Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland. Electronic address: sylvain.perriot@chuv.ch.
² Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland.
³ Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland; Service of Neurology, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland. Electronic address: renaud.du-pasquier@chuv.ch.

Abstract

This protocol describes the generation and characterization of human induced pluripotent stem cells (hiPSCs) from erythroblasts. A key difference with classical protocols is the reprogramming of erythroblasts from a simple blood draw as opposed to fibroblasts/keratinocytes, which requires a biopsy. Moreover, working with erythroblasts ensures that no recombination of the TCR/BCR genes occurs, as opposed to T cells and whole peripheral blood mononuclear cells-based approaches. Last, this approach uses non-integrative episomes ensuring no integration of transgenes into the hiPSCs genome. For complete details on the use and execution of this protocol, please refer to Perriot et al. (2018).

Keywords: Biotechnology and bioengineering; Cell Biology; Cell culture; Cell isolation; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cellular Reprogramming
Erythroblasts
Humans
Induced Pluripotent Stem Cells*
Leukocytes, Mononuclear
Transgenes