Inducible and Deterministic Forward Programming of Human Pluripotent Stem Cells into Neurons, Skeletal Myocytes, and Oligodendrocytes.
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.
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.
Matthias
M
Pawlowski
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.
Daniel
D
Ortmann
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.
Alessandro
A
Bertero
Department of Clinical Neuroscience, University of Cambridge, Cambridge CB2 0QQ, UK.
Joana M
JM
Tavares
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.
Roger A
RA
Pedersen
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.
Ludovic
L
Vallier
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.
Mark R N
MRN
Kotter
D002454:Cell Differentiation; D002460:Cell Line; D065150:Cellular Reprogramming; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D019076:Transgenes; D015854:Up-Regulation
human pluripotent stem cells; neurons; oligodendrocyte progenitor cells; reprogramming; skeletal myocytes