Figure 9

Tracking of transplanted human corneal endothelial cells in vivo. (a) A DIR stained DMEK graft imaged in the viewing chamber pre-implantation. A triangular notch is cut out the circular graft and used to confirm the correct side is up during surgery (yellow arrow). (b) The same graft is imaged immediately after surgery. Linear, parallel wounds associated with graft insertion can be seen following DMEK (red arrow) as can the notch in the transplanted graft (yellow arrow). (c–e) The graft is repeatedly imaged over 28 days. Central areas, previously devoid of cells, become DIR positive suggesting cells migration over DM has occurred (red arrow). The triangular notch fades suggesting cells have migrated off the graft and onto the rabbit DM at this point (yellow arrow). Areas in the periphery of the graft are not DIR fluorescent, suggesting they are devoid of human endothelial cells. (f–j) Images of the rabbit eye show at the time of imaging show that DIR fluorescence is detectable when the anterior chamber contains air (g,h) and that cornea clarity is restored once a stable endothelial monolayer has been established (i,j). OCT images of cornea at various time points following transplantation (k–o). Scale bar (a–j): 1 mm. (p) The triangular notch shows created for orientation identification shows a loss of the straight edge (red dashed line) with apparent migration of endothelial cells off the graft. (q) Images of the same area are acquired after staining with human nuclear antibody (FITC). The presence of human endothelial cells beyond the border of the DMEK graft (red dashed line) is confirmed. Scale bar: 250 µm. (r) Areas that do not stain with DIR are seen in the graft periphery. (s) Cells staining positively with DAPI, but not staining with anti-human nuclear antibody (FITC) can be seen, suggesting these cells are of rabbit origin and that transfer of fluorescence between cells does not occur. Scale bar: 250 µm.