Journal Article

Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells

Zon, Leonard I., Florido, Mary H. C., Prummer, Michael, Adatto, Isaac, Warren, Curtis R., Grainger, Stephanie J., O’Sullivan, John F., Huang, Paul, Challet-Meylan, Ludivine, Xia, Yulei, He, Wei, Graf, Martin, Pan, Wei, Heckel, Tobias, Gerszten, Robert E., Kapp, Friedrich G., Christensen, Klaus, Kling, Dorothee, Jagasia, Ravi, Urich, Eduard, Freskgård, Per-Ola, Iacone, Roberto, Jakob-Roetne, Roland, Thoma, Eva C., Cowan, Chad A., Certa, Ulrich, Sun, Lin, Chaikof, Elliot L., Patsch, Christoph

2015

Overview

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. Cowan and colleagues report a method to generate mature endothelial or vascular smooth muscle cells from human pluripotent stem cells with high efficiency and purity.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13 / 13/100 / 631/136/532/1360 / 631/136/532/2064 / Animals / Biomarkers - metabolism / Blood vessels / Bone Morphogenetic Protein 4 - pharmacology / Cancer Research / Cell Biology / Cell differentiation / Cell Differentiation - drug effects / Cell Line / Cell Lineage - drug effects / Cell research / Coculture Techniques / Developmental Biology / Dose-Response Relationship, Drug / Endothelial Cells - drug effects / Endothelial Cells - enzymology / Endothelial Cells - physiology / Endothelial Cells - transplantation / Endothelium / Gene Expression Profiling - methods / Gene Expression Regulation, Developmental / Glycogen Synthase Kinase 3 - antagonists & inhibitors / Glycogen Synthase Kinase 3 - metabolism / Glycogen Synthase Kinase 3 beta / Growth / 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 / Kinases / Life Sciences / Metabolomics - methods / Mice, Inbred NOD / Mice, SCID / Muscle cells / 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 / Smooth muscle / Stem Cells / Time Factors / Transcription, Genetic / Transfection / Vascular Endothelial Growth Factor A - pharmacology / Wnt Signaling Pathway - drug effects