Fibrin bead assay with VEGF-A stimulation.

<p>Confocal microscopy images of fibrin bead assay.&nbsp;</p> <p>Related article for citation:&nbsp;<a href="https://www.nature.com/articles/s41598-023-33090-6">https://www.nature.com/articles/s41598-023-33090-6</a>&nbsp;&quot;SproutAngio: an open-source bioimage informatics tool for quantitative analysis of sprouting angiogenesis and lumen space&quot;</p> <p>Cells: Human umbilical vein endothelial cells (HUVECs) were isolated,&nbsp;maintained in PromoCell Endothelial Cell Growth Medium (Heidelberg, Germany) with 1X endothelial growth kit addition, and used&nbsp;in passage 2. Human lung pulmonary fibroblasts (hPFs) were purchased from PromoCell,&nbsp;maintained in DMEM with 10% FBS, and used in passage 8.</p> <p>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)</p> <p>We used VEGF-A from R&amp;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.</p> <p>Staining: DAPI staining for nuclei, phalloidin staining for F-actin.</p>