Abstract
The use of human pluripotent stem cells for in vitro disease modelling and clinical applications requires protocols that convert these cells into relevant adult cell types. Here, we report the rapid and efficient differentiation of human pluripotent stem cells into vascular endothelial and smooth muscle cells. We found that GSK3 inhibition and BMP4 treatment rapidly committed pluripotent cells to a mesodermal fate and subsequent exposure to VEGF-A or PDGF-BB resulted in the differentiation of either endothelial or vascular smooth muscle cells, respectively. Both protocols produced mature cells with efficiencies exceeding 80% within six days. On purification to 99% via surface markers, endothelial cells maintained their identity, as assessed by marker gene expression, and showed relevant in vitro and in vivo functionality. Global transcriptional and metabolomic analyses confirmed that the cells closely resembled their in vivo counterparts. Our results suggest that these cells could be used to faithfully model human disease.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Becaplermin
-
Biomarkers / metabolism
-
Bone Morphogenetic Protein 4 / pharmacology
-
Cell Differentiation* / drug effects
-
Cell Line
-
Cell Lineage* / drug effects
-
Coculture Techniques
-
Dose-Response Relationship, Drug
-
Endothelial Cells / drug effects
-
Endothelial Cells / enzymology
-
Endothelial Cells / physiology*
-
Endothelial Cells / transplantation
-
Gene Expression Profiling / methods
-
Gene Expression Regulation, Developmental
-
Glycogen Synthase Kinase 3 / antagonists & inhibitors
-
Glycogen Synthase Kinase 3 / metabolism
-
Glycogen Synthase Kinase 3 beta
-
Human Umbilical Vein Endothelial Cells / physiology
-
Humans
-
Induced Pluripotent Stem Cells / drug effects
-
Induced Pluripotent Stem Cells / enzymology
-
Induced Pluripotent Stem Cells / physiology*
-
Induced Pluripotent Stem Cells / transplantation
-
Metabolomics / methods
-
Mice, Inbred NOD
-
Mice, SCID
-
Muscle, Smooth, Vascular / cytology
-
Muscle, Smooth, Vascular / drug effects
-
Muscle, Smooth, Vascular / enzymology
-
Muscle, Smooth, Vascular / physiology*
-
Muscle, Smooth, Vascular / transplantation
-
Myocytes, Smooth Muscle / drug effects
-
Myocytes, Smooth Muscle / enzymology
-
Myocytes, Smooth Muscle / physiology*
-
Myocytes, Smooth Muscle / transplantation
-
Neovascularization, Physiologic
-
Phenotype
-
Protein Kinase Inhibitors / pharmacology
-
Proto-Oncogene Proteins c-sis / pharmacology
-
Time Factors
-
Transcription, Genetic
-
Transfection
-
Vascular Endothelial Growth Factor A / pharmacology
-
Wnt Signaling Pathway / drug effects
Substances
-
Biomarkers
-
Bone Morphogenetic Protein 4
-
Protein Kinase Inhibitors
-
Proto-Oncogene Proteins c-sis
-
Vascular Endothelial Growth Factor A
-
Becaplermin
-
Glycogen Synthase Kinase 3 beta
-
Glycogen Synthase Kinase 3