Walking the Line: A Fibronectin Fiber-Guided Assay to Probe Early Steps of (Lymph)angiogenesis

• DOI: 10.1371/journal.pone.0145210
• Publication Date: 2015-12-21T18:37:01Z
• Authors: Maria Mitsi; Martin Michael Pe...; Epameinondas Gous...; Alexandra Michael...; Michael Detmar; Viola Vogel
• Keywords: Lymphangiogenesis; Sprouting angiogenesis; Lymphatic Endothelium

Angiogenesis and lymphangiogenesis are highly complex morphogenetic processes, central to many physiological pathological conditions, including development, cancer metastasis, inflammation wound healing. While it is described that extracellular matrix (ECM) fibers involved in the spatiotemporal regulation of angiogenesis, current angiogenesis assays not specifically designed dissect quantify underlying molecular mechanisms how fibrillar nature ECM regulates vessel sprouting. Even less known about role during early stages lymphangiogenesis. To address such questions, we introduced here an vitro (lymph)angiogenesis assay, where used microbeads coated with endothelial cells as simple sprouting sources deposited them on single Fn substrates mimic ECM. The were a transparent substrate, suitable for live microscopic observation ensuing cell outgrowth events at level. Our proof-of-concept studies revealed Fn, compared Fn-coated surfaces, provides far stronger guidance cues cells, independent tested mechanical strains fibers. Additionally, found VEGF-A, but VEGF-C, stimulates collective lymphatic (LEC), while blood vascular (HUVEC) was prominent even absence these angiogenic factors. In addition findings presented here, modularity our assay allows use different or synthetic substrates, well other types, thus expanding range applications biology beyond.