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A. Kubota, M. Akiba, Y. Takayanagi, T. Soma, N. Maeda, C. Kimpui, K. Nishida; Validation of Human and Rabbit Cultured Corneal Epithelial Cell Sheets by Full-Field Optical Coherence Tomography. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2811. doi: https://doi.org/.
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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.
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.
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.
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.
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