title: Endothelial matrix assembly during capillary morphogenesis: insights from chimeric TagRFP-fibronectin matrix.
abstract: Biologically relevant, three-dimensional extracellular matrix is an essential component of in vitro vasculogenesis models. WI-38 fibroblasts assemble a 3D matrix that induces endothelial tubulogenesis, but this model is challenged by fibroblast senescence and the inability to distinguish endothelial cell-derived matrix from matrix made by WI-38 fibroblasts. Matrices produced by hTERT-immortalized WI-38 recapitulated those produced by wild type fibroblasts. ECM fibrils were heavily populated by tenascin-C, fibronectin, and type VI collagen. Nearly half of the total type I collagen, but only a small fraction of the type IV collagen, were incorporated into ECM. Stable hTERT-WI-38 transfectants expressing TagRFP-fibronectin incorporated TagRFP into ~90% of the fibronectin in 3D matrices. TagRFP-fibronectin colocalized with tenascin-C and with type I collagen in a pattern that was similar to that seen in matrices from wild type WI-38. Human Umbilical Vein Endothelial Cells (HUVEC) formed 3D adhesions and tubes on WI38-hTERT-TagRFP-FN-derived matrices, and the TagRFP-fibronectin component of this new 3D human fibroblast matrix model facilitated the demonstration of concentrated membrane type 1 metalloprotease and new HUVEC FN and collagen type IV fibrils during EC tubulogenesis. These findings indicate that WI-38-hTERT- and WI-38-hTERT-TagRFP-FN-derived matrices provide platforms for the definition of new matrix assembly and remodeling events during vasculogenesis.
authors: Anesthesiology and Critical Care Medicine (FC, LR), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA (CAL)Cell Biology (LR), Johns Hopkins Medical Institutions, Baltimore, MDBiomedical Engineering (LR), Johns Hopkins Medical Institutions, Baltimore, MDPediatrics (LR), Johns Hopkins Medical Institutions, Baltimore, MDCenter for Cell Dynamics (LR), Johns Hopkins Medical Institutions, Baltimore, MDGraduate Program in Cellular and Molecular Medicine (VN), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Molecular and Cell Biology, The Weizmann Institute of Science, Rehovot, Israel (VR).
authors: Fumin
authors: F
authors: Chang
authors: Anesthesiology and Critical Care Medicine (FC, LR), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA (CAL)Cell Biology (LR), Johns Hopkins Medical Institutions, Baltimore, MDBiomedical Engineering (LR), Johns Hopkins Medical Institutions, Baltimore, MDPediatrics (LR), Johns Hopkins Medical Institutions, Baltimore, MDCenter for Cell Dynamics (LR), Johns Hopkins Medical Institutions, Baltimore, MDGraduate Program in Cellular and Molecular Medicine (VN), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Molecular and Cell Biology, The Weizmann Institute of Science, Rehovot, Israel (VR).
authors: Christopher A
authors: CA
authors: Lemmon
authors: Anesthesiology and Critical Care Medicine (FC, LR), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA (CAL)Cell Biology (LR), Johns Hopkins Medical Institutions, Baltimore, MDBiomedical Engineering (LR), Johns Hopkins Medical Institutions, Baltimore, MDPediatrics (LR), Johns Hopkins Medical Institutions, Baltimore, MDCenter for Cell Dynamics (LR), Johns Hopkins Medical Institutions, Baltimore, MDGraduate Program in Cellular and Molecular Medicine (VN), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Molecular and Cell Biology, The Weizmann Institute of Science, Rehovot, Israel (VR).
authors: Voraphoj
authors: V
authors: Nilaratanakul
authors: Anesthesiology and Critical Care Medicine (FC, LR), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA (CAL)Cell Biology (LR), Johns Hopkins Medical Institutions, Baltimore, MDBiomedical Engineering (LR), Johns Hopkins Medical Institutions, Baltimore, MDPediatrics (LR), Johns Hopkins Medical Institutions, Baltimore, MDCenter for Cell Dynamics (LR), Johns Hopkins Medical Institutions, Baltimore, MDGraduate Program in Cellular and Molecular Medicine (VN), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Molecular and Cell Biology, The Weizmann Institute of Science, Rehovot, Israel (VR).
authors: Varda
authors: V
authors: Rotter
authors: Anesthesiology and Critical Care Medicine (FC, LR), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA (CAL)Cell Biology (LR), Johns Hopkins Medical Institutions, Baltimore, MDBiomedical Engineering (LR), Johns Hopkins Medical Institutions, Baltimore, MDPediatrics (LR), Johns Hopkins Medical Institutions, Baltimore, MDCenter for Cell Dynamics (LR), Johns Hopkins Medical Institutions, Baltimore, MDGraduate Program in Cellular and Molecular Medicine (VN), Johns Hopkins Medical Institutions, Baltimore, MDDepartment of Molecular and Cell Biology, The Weizmann Institute of Science, Rehovot, Israel (VR) LRomer@jhmi.edu.
authors: Lewis
authors: L
authors: Romer
mesh_terms: D002196:Capillaries / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism*; D000097573:Red Fluorescent Protein
chemical_list: D005353:Fibronectins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: 3D-matrix; endothelium; fibronectin; vasculogenesis