title: In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing.
abstract: In vitro angiogenesis assays are commonly used to assess pro- or anti-angiogenic drug properties. Extracellular matrix (ECM) substitutes such as Matrigel and collagen gel became very popular in in vitro 3D angiogenesis assays as they enable tubule formation by endothelial cells from culture or aortic rings. However, these assays are usually used with a single cell type, lacking the complex cellular interactions occurring during angiogenesis. Here, we report a novel angiogenesis assay using egg white as ECM substitute. We found that, similar to Matrigel, egg white elicited prevascular network formation by endothelial and/or smooth muscle cell coculture. This matrix was suitable for various cells from human, mouse, and rat origin. It is compatible with aortic ring assay and also enables vascular and tumor cell coculture. Through simple labeling (DAPI, Hoechst 33258), cell location and resulting prevascular network formation can easily be quantified. Cell transfection with green fluorescent protein improved whole cell visualization and 3D structure characterization. Finally, egg-based assay dedicated to angiogenesis studies represents a reliable and cost-effective way to produce and analyze data regarding drug effects on vascular cells.
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Yoanne
authors.initials: Y
authors.lastname: Mousseau
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Mollard
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Hao
authors.initials: H
authors.lastname: Qiu
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Richard
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Cazal
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Angélique
authors.initials: A
authors.lastname: Nizou
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Vedrenne
authors.affiliation: INSERM U 643, Nantes, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Rémi
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Yasser
authors.initials: Y
authors.lastname: Baaj
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Fourcade
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Funalot
authors.affiliation: Department of Biochemistry and Molecular Genetics, EA 6063, CHU Dupuytren, Limoges, France.
authors.forename: Franck G
authors.initials: FG
authors.lastname: Sturtz
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001013:Aorta, Thoracic / Q000166:cytology; D006690:Bisbenzimidazole; D018920:Coculture Techniques / Q000379:methods; D003094:Collagen; D004338:Drug Combinations; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004529:Egg White; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D005456:Fluorescent Dyes; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007211:Indoles; D007797:Laminin; D051379:Mice; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011509:Proteoglycans; D051381:Rats; D013045:Species Specificity
chemical_list: D004338:Drug Combinations; D005456:Fluorescent Dyes; D007211:Indoles; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; C007293:DAPI; D003094:Collagen; D006690:Bisbenzimidazole