Development of endothelial cell lines from embryonic stem cells: A tool for studying genetically manipulated endothelial cells in vitro.
Totipotent embryonic stem cells can be induced to differentiate to endothelium in vitro. This may be a useful tool for obtaining cultures of genetically manipulated endothelial cells because embryonic stem cells are relatively easy to transfect and are commonly used for gene inactivation experiments in mice. However, embryonic stem cell-derived endothelial cells could not be easily separated from embryoid bodies and maintained in culture. In this study, we describe the isolation and characterization of immortalized endothelial cell lines obtained from embryonic stem cells differentiated in vitro. The cell lines were analyzed for expression of endothelial cell markers, including growth factor receptors and adhesion molecules, and compared with endothelial cells obtained from the yolk sac, the embryo proper, or the heart microcirculation of the adult. We propose that this approach may be useful for obtaining endothelial cells carrying gene mutations that are lethal at very early stages of development.
Balconi
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Istituto di Ricerche Farmacologiche Mario Negri (G.B., R.S., E.D.), Milan, Italy. balconi@irfmn.mnegri.it
Spagnuolo
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Dejana
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D000818:Animals; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D051929:CD146 Antigen; D015815:Cell Adhesion Molecules / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000032:analysis; D016228:Endothelial Growth Factors / Q000032:analysis; D004730:Endothelium, Vascular / Q000166:cytology* / Q000196:embryology*; D005455:Fluorescent Antibody Technique; D005821:Genetic Techniques; D016023:Integrins / Q000032:analysis; D062726:Junctional Adhesion Molecules; D008222:Lymphokines / Q000032:analysis; D008562:Membrane Glycoproteins; D051379:Mice; D018919:Neovascularization, Physiologic; D019006:Neural Cell Adhesion Molecules; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*; D015534:Trans-Activators; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D015017:Yolk Sac / Q000166:cytology; D051176:beta Catenin
D015703:Antigens, CD; D018952:Antigens, CD34; D000954:Antigens, Surface; D015415:Biomarkers; D051929:CD146 Antigen; C495267:CTNNB1 protein, mouse; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D016228:Endothelial Growth Factors; D016023:Integrins; D062726:Junctional Adhesion Molecules; D008222:Lymphokines; D008562:Membrane Glycoproteins; D019006:Neural Cell Adhesion Molecules; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D015534:Trans-Activators; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D051176:beta Catenin