Validation of Human and Rabbit Cultured Corneal Epithelial Cell Sheets by Full-Field Optical Coherence Tomography

Abstract

Purpose: : To observe the morphology and thickness of cultured human and rabbit corneal epithelial cell sheets alive and non-contact by use of a prototype full-field optical coherence tomography (FF-OCT) setup that offers ultrahigh (<1µm), three dimensional optical sectioning capability.

Methods: : Human and rabbit corneal epithelial cell sheets were prepared using the method that we previously reported. For the preparation of lethally treated National Institutes of Health (NIH)/3T3 feeder layers, subconfluent NIH/3T3 fibroblasts were incubated with 16 _g/mL of mitomycin C (MMC) for 2 hours at 37°C, and then trypsinized and seeded onto the temperature-responsive culture dishes at a density of 2x10⁴ cells/cm². Both the cultured sheets and the nucleated rabbit corneal epithelium were imaged by using the FF-OCT setup.

Results: : Basal cells, wing cells, and superficial cells in both enucleated rabbit cornea and cultured corneal cell sheets were observed at a cell level with the FF-OCT. The growth process of epithelial cell sheet could be monitored using the FF-OCT setup, and the thickness of the cultured epithelial sheet was measured quantitatively.

Conclusions: : FF-OCT is feasible for a cellular level observation of cultured epithelial cell sheet alive and non-contact. It may provide a useful mean for the validation of cultured epithelial cell sheets in regenerative medicine.

Keywords: cornea: clinical science • cornea: epithelium • imaging/image analysis: clinical