Figure 10.

Ciliary zonule-like protein matrix promotes migration of immune cells. (A) Phase contrast image of primary cultures of human non-pigmented ciliary epithelial cells (HNPCEpiCs), the ciliary body cell type that produces the proteins that comprise the ciliary zonules (CZ). (B to D) Confocal microscopy imaging of HNPCEpiC-conditioned matrix substrates immunolabeled for (B) fibronectin, (C) fibrillin-1, or (D) tenascin-C. (E, F) Representative tracks of cells traced with Imaris software from 24-h time-lapse imaging of RAW 264.7 cells migrating on (E) HNPCEpiC-conditioned substrates and (F) tissue culture substrates. (Ei, Fi) phase contrast images of the position of cells at time 0 of the time-lapse study overlaid with the track of the cells with purple denoting the time 0 start site and red the cells’ position at 24 h. (Eii, Fii) Images showing the traced cell paths alone. (G to J) Bar graphs quantifying Imaris image analytics of 126 cells tracked on HNPCEpiC-conditioned substrates as compared to 114 cells tracked on tissue culture plastic to determine (G) track length, (H) track displacement, (I) track speed, and (J) track straightness. While the immune cells had a similar speed and overall track length, there were significant differences in how far the cells moved and their ability to move in a directional manner, with the HNPCEpiC-produced matrix substrate promoting directional immune cell migration. (A color version of this figure is available in the online journal.)