Inducible and Deterministic Forward Programming of Human Pluripotent Stem Cells into Neurons, Skeletal Myocytes, and Oligodendrocytes

Matthias Pawlowski; Daniel Ortmann; Alessandro Bertero; Joana M Tavares; Roger A Pedersen; Ludovic Vallier; Mark R N Kotter

doi:10.1016/j.stemcr.2017.02.016

Inducible and Deterministic Forward Programming of Human Pluripotent Stem Cells into Neurons, Skeletal Myocytes, and Oligodendrocytes

Stem Cell Reports. 2017 Apr 11;8(4):803-812. doi: 10.1016/j.stemcr.2017.02.016. Epub 2017 Mar 23.

Authors

Matthias Pawlowski¹, Daniel Ortmann², Alessandro Bertero², Joana M Tavares³, Roger A Pedersen⁴, Ludovic Vallier⁵, Mark R N Kotter⁶

Affiliations

¹ Anne McLaren Laboratory, Wellcome Trust-MRC Stem Cell Institute, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Clinical Neuroscience, University of Cambridge, Cambridge CB2 0QQ, UK. Electronic address: pawlowsk@uni-muenster.de.
² Anne McLaren Laboratory, Wellcome Trust-MRC Stem Cell Institute, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Surgery, University of Cambridge, Cambridge CB2 0QQ, UK.
³ Department of Clinical Neuroscience, University of Cambridge, Cambridge CB2 0QQ, UK.
⁴ Anne McLaren Laboratory, Wellcome Trust-MRC Stem Cell Institute, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Paediatrics, University of Cambridge, Cambridge, CB2 0QQ, UK.
⁵ Anne McLaren Laboratory, Wellcome Trust-MRC Stem Cell Institute, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Surgery, University of Cambridge, Cambridge CB2 0QQ, UK; Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
⁶ Anne McLaren Laboratory, Wellcome Trust-MRC Stem Cell Institute, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Clinical Neuroscience, University of Cambridge, Cambridge CB2 0QQ, UK. Electronic address: mrk25@cam.ac.uk.

Abstract

The isolation or in vitro derivation of many human cell types remains challenging and inefficient. Direct conversion of human pluripotent stem cells (hPSCs) by forced expression of transcription factors provides a potential alternative. However, deficient inducible gene expression in hPSCs has compromised efficiencies of forward programming approaches. We have systematically optimized inducible gene expression in hPSCs using a dual genomic safe harbor gene-targeting strategy. This approach provides a powerful platform for the generation of human cell types by forward programming. We report robust and deterministic reprogramming of hPSCs into neurons and functional skeletal myocytes. Finally, we present a forward programming strategy for rapid and highly efficient generation of human oligodendrocytes.

Keywords: human pluripotent stem cells; neurons; oligodendrocyte progenitor cells; reprogramming; skeletal myocytes.

Publication types

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

MeSH terms

Cell Differentiation*
Cell Line
Cellular Reprogramming
Gene Expression
Gene Targeting / methods*
Humans
Muscle Development
Muscle Fibers, Skeletal / cytology*
Muscle Fibers, Skeletal / metabolism
Neurogenesis
Neurons / cytology*
Neurons / metabolism
Oligodendroglia / cytology*
Oligodendroglia / metabolism
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / metabolism
Transgenes
Up-Regulation

Abstract

Publication types

MeSH terms

Grants and funding