Fig. 5

miR-184 suppresses growth and migration of human primary choroidal endothelial cells (hCEC) A Schematic of isolation of endothelial cells from fresh donor eyes using CD31( +) magnetic beads and culture and assay of miR-184 mimic and inhibitor transfected cells. B 3D Tube formation assay using miR-184 mimic/inhibitor transfected hCEC (primary human choroidal endothelial cell) at 4 h after seeding. Images were skeletonized and measured for analysis. Scrambled negative controls for mimic and inhibitor are indicated as N.C. C, D Total C loops and D branching points were measured for tube formation property parameters at 4 h. Note a significant decrement of total loops and branching points at miR-184 mimic transfected cells. E Wound healing assay using miR-184 mimic/inhibitor-transfected hCEC. The wound space was measured at 2 h after scratching. F, G Measurement of the wound space area. F miR-184 mimic transfected cells showed reduced wound closure, while G no significant difference was observed in the miR-184 inhibitor transfected cells. H Schematic of sprouting angiogenesis assay using microfluidics chip. 20 ng/mL of VEGF gradient was generated to induce endothelial cell sprouting. I Sprouting angiogenesis assay using miR-184 mimic/inhibitor transfected hCEC. Cells were analyzed at 48 h after generating a VEGF gradient. J The number of tip cells per mm was measured and analyzed. miR-184 mimic transfected cells express significantly low tip cells. All assays were performed with three replicative cultures. Graph values are represented as mean ± standard deviation. Statistical significance indicated as *P < 0.05, **P < 0.01, (all n = 3), B, C Scale bar = 1 mm. E–I Scale bar = 20 μm