title: Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells.
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.
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Christoph
authors.initials: C
authors.lastname: Patsch
authors.affiliation: Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA.
authors.forename: Ludivine
authors.initials: L
authors.lastname: Challet-Meylan
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Eva C
authors.initials: EC
authors.lastname: Thoma
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Eduard
authors.initials: E
authors.lastname: Urich
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Tobias
authors.initials: T
authors.lastname: Heckel
authors.affiliation: Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: John F
authors.initials: JF
authors.lastname: O'Sullivan
authors.affiliation: 1] Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, 110 Francis Street, Suite 9F Boston, Massachusetts 02115, USA [2] The Wyss Institute of Biologically Inspired Engineering of Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Grainger
authors.affiliation: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Friedrich G
authors.initials: FG
authors.lastname: Kapp
authors.affiliation: Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Sun
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Christensen
authors.affiliation: Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA.
authors.forename: Yulei
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA.
authors.forename: Mary H C
authors.initials: MH
authors.lastname: Florido
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Wei
authors.initials: W
authors.lastname: Pan
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Prummer
authors.affiliation: Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA.
authors.forename: Curtis R
authors.initials: CR
authors.lastname: Warren
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Roland
authors.initials: R
authors.lastname: Jakob-Roetne
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Certa
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Ravi
authors.initials: R
authors.lastname: Jagasia
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Per-Ola
authors.initials: PO
authors.lastname: Freskgård
authors.affiliation: Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: Isaac
authors.initials: I
authors.lastname: Adatto
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Kling
authors.affiliation: Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Huang
authors.affiliation: 1] Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA [2] Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Leonard I
authors.initials: LI
authors.lastname: Zon
authors.affiliation: 1] Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, 110 Francis Street, Suite 9F Boston, Massachusetts 02115, USA [2] The Wyss Institute of Biologically Inspired Engineering of Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Chaikof
authors.affiliation: Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Gerszten
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Graf
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Roberto
authors.initials: R
authors.lastname: Iacone
authors.affiliation: 1] Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Massachusetts 02138, USA [2] Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA [3] Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Chad A
authors.initials: CA
authors.lastname: Cowan
mesh_terms: D000818:Animals; D000077214:Becaplermin; D015415:Biomarkers / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology / Q000502:physiology* / Q000637:transplantation; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000201:enzymology / Q000502:physiology* / Q000637:transplantation; D055432:Metabolomics / Q000379:methods; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000502:physiology* / Q000637:transplantation; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000201:enzymology / Q000502:physiology* / Q000637:transplantation; D018919:Neovascularization, Physiologic; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4; D047428:Protein Kinase Inhibitors; D020574:Proto-Oncogene Proteins c-sis; D042461:Vascular Endothelial Growth Factor A; D000077214:Becaplermin; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3