title: Three-Dimensional Vascular Network Assembly From Diabetic Patient-Derived Induced Pluripotent Stem Cells.
abstract: OBJECTIVE
  In diabetics, hyperglycemia results in deficient endothelial progenitors and cells, leading to cardiovascular complications. We aim to engineer 3-dimensional (3D) vascular networks in synthetic hydrogels from type 1 diabetes mellitus (T1D) patient-derived human-induced pluripotent stem cells (hiPSCs), to serve as a transformative autologous vascular therapy for diabetic patients.
  
  
  RESULTS
  We validated and optimized an adherent, feeder-free differentiation procedure to derive early vascular cells (EVCs) with high portions of vascular endothelial cadherin-positive cells from hiPSCs. We demonstrate similar differentiation efficiency from hiPSCs derived from healthy donor and patients with T1D. T1D-hiPSC-derived vascular endothelial cadherin-positive cells can mature to functional endothelial cells-expressing mature markers: von Willebrand factor and endothelial nitric oxide synthase are capable of lectin binding and acetylated low-density lipoprotein uptake, form cords in Matrigel and respond to tumor necrosis factor-α. When embedded in engineered hyaluronic acid hydrogels, T1D-EVCs undergo morphogenesis and assemble into 3D networks. When encapsulated in a novel hypoxia-inducible hydrogel, T1D-EVCs respond to low oxygen and form 3D networks. As xenografts, T1D-EVCs incorporate into developing zebrafish vasculature.
  
  
  CONCLUSIONS
  Using our robust protocol, we can direct efficient differentiation of T1D-hiPSC to EVCs. Early endothelial cells derived from T1D-hiPSC are functional when mature. T1D-EVCs self-assembled into 3D networks when embedded in hyaluronic acid and hypoxia-inducible hydrogels. The capability of T1D-EVCs to assemble into 3D networks in engineered matrices and to respond to a hypoxic microenvironment is a significant advancement for autologous vascular therapy in diabetic patients and has broad importance for tissue engineering.
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Xin Yi
authors.initials: XY
authors.lastname: Chan
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Black
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Kayla
authors.initials: K
authors.lastname: Dickerman
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Joseph
authors.initials: J
authors.lastname: Federico
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Lévesque
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Jeff
authors.initials: J
authors.lastname: Mumm
authors.affiliation: From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Sharon
authors.initials: S
authors.lastname: Gerecht
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D016022:Case-Control Studies; D002454:Cell Differentiation; D015687:Cell Hypoxia; D002460:Cell Line; D002469:Cell Separation; D048430:Cell Shape; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000473:pathology*; D066026:Endothelial Progenitor Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D064593:Heterografts; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry; D020100:Hydrogels; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D008077:Lipoproteins, LDL / Q000378:metabolism; D009389:Neovascularization, Pathologic; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D010641:Phenotype; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D049452:Green Fluorescent Proteins; D006820:Hyaluronic Acid; D020100:Hydrogels; D008077:Lipoproteins, LDL; D052250:Nitric Oxide Synthase Type III; D014409:Tumor Necrosis Factor-alpha; D014841:von Willebrand Factor; D000099152:Cadherin 5; C032629:acetyl-LDL; C496314:NOS3 protein, human
keywords: diabetes mellitus; endothelial cells; hydrogels; induced pluripotent stem cells; vascular networks