Transient expression of OCT4 is sufficient to allow human keratinocytes to change their differentiation pathway.
In this study, we describe a simple system in which human keratinocytes can be redirected to an alternative differentiation pathway. We transiently transfected freshly isolated human skin keratinocytes with the single transcription factor OCT4. Within 2 days these cells displayed expression of endogenous embryonic genes and showed reduced genomic methylation. More importantly, these cells could be specifically converted into neuronal and contractile mesenchymal cell types. Redirected differentiation was confirmed by expression of neuronal and mesenchymal cell mRNA and protein, and through a functional assay in which the newly differentiated mesenchymal cells contracted collagen gels as efficiently as authentic myofibroblasts. Thus, to generate patient-specific cells for therapeutic purposes, it may not be necessary to completely reprogram somatic cells into induced pluripotent stem cells before altering their differentiation and grafting them into new tissues.
Racila
D
D
Department of Anatomy and Cell Biology, University of Iowa Carver College of Medicine, Iowa City, IA, USA.
Winter
M
M
Said
M
M
Tomanek-Chalkley
A
A
Wiechert
S
S
Eckert
R L
RL
Bickenbach
J R
JR
D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D012313:RNA / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
D017931:DNA Primers; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012313:RNA