Effects of citicoline on wound recovery, spheroid sprouting and tube-like structure formation in Matrigel. A) a significant increase in the number of migrating cells from a scratch wound was seen in hCMEC 24 h after treatment with FGF-2 (positive control; ii) and citicoline (10 μM; p = 0.015; iii). The bar chart represents the mean numbers of migrating cells from triplicate coverslips of control untreated cells, FGF-2 positive control (25 ng/ml), and citicoline treated cells (10 μM). B) Shows that addition of citicoline (10 μM) significantly increased the formation and intensity of spheroids as well as migration of cells outwards from the central mass after 7 days incubation (P = 0.005; iii). The bar chart represents the mean sprout length of control spheroids after 7 days, FGF-2 (25 ng/ml), and Citicoline 10 μM. Values are expressed in μM and at least 10 spheroids from each condition were analyzed. C) When cells were cultured in Matrigel™ in the presence of citicoline for 24 h, a significant increase in the formation of tube-like structures was observed compared with control cells (p = 0.010; iii). The bar graphs and photomicrographs shows control cells, cells treated with FGF-2 (25 ng/ml), cells treated with citicoline (10 μg/ml). The data illustrates the significant increase in the numbers of closed-loop areas in the presence of citicoline (p = 0.010). Five areas from each well and 3 wells per treatment were analyzed.