Fig. 7
Network map of potential proteins/pathways involved in scCM-induced effects. (a) Schematic overview of angiogenesis process and in vitro assays proposed for evaluating EPC-secretome effects. Human CD34+-derived cord-blood hematopoietic endothelial cells (CD34+-ECs) were used to study angiogenesis (proliferation, cell migration, and tubulogenesis). To investigate mechanisms involved in vascular maturation, CD34+-EC were seeded on the surface of Matrigelâ„¢-coated Transwell inserts and cultivated in monocultures (CMECs). Finally, to study the effect of EPC-secretome on brain capillaries, we used an in vitro BBB model which consisted in seeding CD34+-ECs on Matrigelâ„¢-coated Transwell inserts and co-cultivating them with human brain pericytes, which enabled CD34+-ECs to acquire a brain-like endothelial cell phenotype (BLECs). (b) Proteins present in scCM were converted in gene identifiers and were then grouped according to their biological function/pathway (metabolic pathways, growth factors, inflammatory response, basal membrane (BM) regulation, cytoskeletal regulation, and integrin pathway). An interaction network was prepared using the Cytoscape software (version 3.8.2, released 2020.10.24). The regulation of MAPK activity might play an essential role in the scCM-induced angiogenesis in human primary ECs. In parallel, scCM promotes vascular maturation of newly formed vessels and protects the BBB integrity under ischemic and inflammatory conditions