The Classical Matrigel Angiogenesis Model Revisited: Key Regulators and Comparative Analysis with 2D/3D Co-Culture...

Capillary-like network (CLN) formation on Matrigel is a classical model for assessing the angiogenic properties of cells, though the underlying mechanisms vary across cell types. We identified two distinct CLN formation patterns and evaluated how each type characterizes the angiogenic potential of cells. Only endothelial cells (ECs) capable of elongating during CLN assembly on Matrigel exhibited angiogenic activity in other in vitro models (2D and 3D). Transcriptional profiling of ECs in a 2D co-culture with MSCs showed that CLN formation coincided with the upregulation of genes encoding tight junction proteins, Notch pathway components, and syndecan-2. Stromal cells are also capable of forming CLNs on Matrigel and in 3D model, underscoring their physiological role in tissue remodeling and homeostasis through network assembly.