Abstract
Purpose :
The corneal epithelium (CE) often sustains damage from trauma due to its position as the interface between the cornea and the external environment. A properly structured cornea is required for clear vision, so ensuring the CE heals correctly is crucial. We have previously shown that c-Met activity drives corneal epithelial cell migration and is limited by its desensitization after HGF stimulation. The goal of this research is to determine if the findings in immortalized cells can be extended to more sophisticated models like primary human cells and intact eyes.
Methods :
Primary human corneal epithelial cells were cultured from corneas that could not be used for transplantation from consenting donors (The Eye Bank of Kentucky). Primary corneal epithelial cells were treated with 50 ng/ml HGF for 0-240 minutes. c-Met activity was measured by immunoblotting using phosphospecific c-Met antibodies. Corneal epithelial re-epithelialization was measured by monitoring the size of a 1.5 mm debridement wound in the central cornea of adult female C57/Bl6 mouse. The wound was documented by fluorescein staining and imaged by fluorescence microscopy 16 hours after wounding. Wound sizes were quantified using Image J.
Results :
We found that primary corneal epithelial cells respond to HGF in a similar manner to immortalized cells. in vivo studies indicate that HGF stimulates re-epithelialization of debrided mouse corneas.
Conclusions :
These findings put us on track to test whether desensitization of c-Met receptor limits c-Met-mediated re-epithelialization. Our next steps are to establish a knockout mouse model to test desensitization in vivo.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.