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Updated: Apr 20, 2026

In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing
Published on: May 11, 2021
Three-dimensional in vitro assay of endothelial cell invasion and capillary tube morphogenesis
Laura di Blasio1, Federico Bussolino, Luca Primo
1Department of Oncology, Institute for Cancer Research and Treatment of Candiolo, University of Torino, Torino, Italy.
Abstract:
In vitro assays with endothelial cells (EC) cultured on three-dimensional gel recapitulate several aspects of vascular morphogenesis and pathological angiogenesis. The two most used in vitro assays of vascular morphogenesis are the tube formation on extracellular matrix gel and the sprouting from EC spheroids. Tube formation assay measures the ability of EC, plated on gel derived from reconstituted basement membrane, to form capillary-like structures. Sprouting assay is based on spheroids of EC, embedded in collagen gel and stimulated with angiogenic factors, which originate a complex network of capillary-like structures invading the gel. Both these assays can be exploited for antiangiogenic drug screening and gene function analysis during vascular morphogenesis.

