Fig. 7
CCR5-overexpressing MSCs possess increased migrative capacity towards CCL5 in vitro and to EAU retinas in vivo. (A) Two-dimensional wound healing test was performed to detect the migration of MSCtdTomato and MSCCCR5 cells toward hCCL5. Shown are phase-contrast images of the cell culture at the indicated timepoints following the removal of culture-insert. (B) Transwell migration assay was performed to detect the migration of MSCtdTomato and MSCCCR5 cells toward hCCL5. MSCs migrated to the lower chamber surface were stained with crystal violet. (C) Quantification of gap closure in the wound healing test showed a significant increase in wound closure rate for MSCCCR5 cells compared to MSCtdTomato cells. Data are presented as mean ± SD (n = 3 individual samples per group). *p < 0.01. (D) Quantification of migrated cells in the transwell migration assay showed a significant increase for MSCCCR5 cells compared to MSCtdTomato cells. Data are presented as mean ± SD (n = 3 individual samples per group). *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significance. (E) Schematic diagram of EAU induction and MSC transplantation into EAU mice. (F) Confocal images of transplanted tdTomato-positive MSCs located in the retina at 13 days post-injection. (G) Quantification of tdTomato-positive cells in retinas from mice transplanted with MSCCCR5 or MSCtdTomato cells. Data are presented as mean ± SD (n = 7–10 retinas per group). *p < 0.05. d.p.i.: day(s) postimmunization; GCL: ganglion cell layer; INL: inner nuclear layer; IRBP: interphotoreceptor retinoid-binding protein; ONL: outer nuclear layer; PTX: pertussis toxin. Scale bar: (A, B) 100 μm, (F), 20 μm