Reconstruction of a rabbit corneal epithelium on a lyophilized amniotic membrane using tilting air-liquid interface culture followed by tilting submerged culture |
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Authors: | Jae-Il Ahn In-Keun Jang Young-Kwon Seo Hee-Hoon Yoon Yeon-Ho Shin Jae-Chan Kim Kye-Yong Song Eun-Kyung Yang Jung-Keug Park |
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Institution: | (1) Department of Chemical and Biochemical Engineering, Dongguk University, Seoul, 100-715, Korea;(2) Department of Ophthalmology, Chung-Ang University Hospital, Seoul, 140-757, Korea;(3) Department of Pathology, College of Medicine, Chung-Ang University, Seoul, 156-755, Korea;(4) R&D Center, Bioland Ltd., Cheonan, 330-863, Korea |
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Abstract: | Herein, we reconstructed a rabbit corneal epithelium on a lyophilized amniotic membrane (LAM) using a modified version of
two Teflon rings (the Ahn’s supporter). We compared the corneal epithelial cells we had differentiated in vitro using air-liquid interface (6 days, 12 days) and submerged (6 days, 12 days) cultures and followed a six-day tilting dynamic
air-liquid interface culture with a six-day tilting submerged culture. We characterized the reconstructed corneal epithelium
using digital photography, histological imaging, and transmission electron microscopy. The reconstructed corneal epithelium
created under air-liquid interface culture exhibited a healthier basal corneal epithelial layer than that created under submerged
culture. The reconstructed corneal epithelium on the LAM that was produced using the tilting dymanic culture exhibited a healthy
basal layer. We therefore proposed that tilting submerged culture not only supplied nutrients from the medium to the corneal
epithelial cells on the LAM, but it also removed the horny layer in the upper part of the reconstructed corneal epithelium,
presumably by mimicking the effects of blinking. This study demonstrated that corneal epithelium reconstruction on a LAM using
a tilting submerged culture after a tilting air-liquid interface culture may be useful not only for allogeneic or autologous
transplantation, but also for in vitro toxicological test kits. |
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Keywords: | lyophilized amniotic membrane corneal epithelium blinking effects horny layer |
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