Protocol to study juxtacrine signaling in human vascular cells using a fluorescence labeling co-culture approach

Endothelial cells interact closely with adjacent endothelial cells and pericytes. The intercellular crosstalk between these cells is essential for angiogenesis, maturation, and vascular homeostasis. This protocol outlines a method to study these interactions using a co-culture system. We describe steps for transducing/transfecting, labeling, and co-culturing human vascular endothelial cells and pericytes. We then detail procedures for immunofluorescence staining and microscopy imaging. This protocol enables molecular in vitro investigations of juxtacrine signaling pathways and gene expression. For complete details on the use and execution of this protocol, please refer to Herrera et al.1.