Angiogenesis Invasion Assay to Study Endothelial Cell Invasion and Sprouting Behavior.

Autor:	Dong Y; University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France., Alonso F; University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France., Jahjah T; University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France., Fremaux I; University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France., Génot E; University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France. elisabeth.genot@inserm.fr.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2608, pp. 345-364.
DOI:	10.1007/978-1-0716-2887-4_20
Abstrakt:	Angiogenesis is the formation of new blood vessels from the existing vasculature. It is a fundamental process in developmental biology but also a pathological event that initiates or aggravates many diseases. In this complex multistep process, endothelial cells are activated by angiogenic stimuli; undergo specialization in response to VEGF/Notch signaling; degrade the basement membrane of the parent vessel; sprout, migrate, and proliferate to form capillary tubes that branch; and ultimately anastomose with adjacent vessels. Here we describe an assay that mimics the invasion step in vitro. Human microvascular endothelial cells are confronted by a VEGF-enriched basement membrane material in a three-dimensional environment that promotes endothelial cell sprouting, tube formation, and anastomosis. After a few hours, endothelial cells have become tip cells, and vascular sprouts can be observed by phase contrast, fluorescence, or time-lapse microscopy. Sprouting endothelial cells express tip cell markers, display podosomes and filopodia, and exhibit cell dynamics similar to those of angiogenic endothelial cells in vivo. This model provides a system that can be manipulated genetically to study physiological or pathological angiogenesis and that can be used to screen compounds for pro-/anti-angiogenic properties. In this chapter, we describe the key steps in setting up this assay. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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