Fibrin bead assay with VEGF-A stimulation.

Confocal microscopy images of fibrin bead assay. Related article for citation: https://www.nature.com/articles/s41598-023-33090-6 "SproutAngio: an open-source bioimage informatics tool for quantitative analysis of sprouting angiogenesis and lumen space" Cells: Human umbilical vein endothelial cells (HUVECs) were isolated, maintained in PromoCell Endothelial Cell Growth Medium (Heidelberg, Germany) with 1X endothelial growth kit addition, and used in passage 2. Human lung pulmonary fibroblasts (hPFs) were purchased from PromoCell, maintained in DMEM with 10% FBS, and used in passage 8. Experimental groups with VEGF-A concentrations: Group1 (no VEGFA), Group2 (1 ng/ml VEGFA), Group3 (10 ng/ml VEGFA), Group4 (20 ng/ml VEGFA), Group5 (50 ng/ml VEGFA) We used VEGF-A from R&D Systems, Minneapolis, MN for the stimulation. Media change and VEGF-A stimulation were done on days 0,2,4,6. Assay fixation was on day 7. Staining: DAPI staining for nuclei, phalloidin staining for F-actin.