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A new 3D reconstituted human corneal epithelium model as an alternative method for the eye irritation test
Authors:Kyoung-Mi JungSu-Hyon Lee  Yang-Hwan RyuWon-Hee Jang  Haeng-Sun JungJu-Hee Han  Seung-Hyeok SeokJae-Hak Park  Youngsook SonYoung-Ho Park  Kyung-Min Lim
Affiliation:a Amorepacific Co. R&D Center, Yongin 446-729, Republic of Korea
b Modern Cell and Tissue Technologies Inc., Seoul 139-743, Republic of Korea
c College of Veterinary Medicine, Seoul National University, Seoul 151-742, Republic of Korea
d College of Life Science, Kyung Hee University, Yongin 441-706, Republic of Korea
Abstract:Many efforts are being made to develop new alternative in vitro test methods for the eye irritation test. Here we report a new reconstructed human corneal epithelial model (MCTT HCE model) prepared from primary-cultured human limbal epithelial cells as a new alternative in vitro eye irritation test method. In histological and immunohistochemical observation, MCTT HCE model displayed a morphology and biomarker expressions similar to intact human cornea. Moreover, the barrier function was well preserved as measured by high transepithelial electrical resistance, effective time-50 for Triton X-100, and corneal thickness. To employ the model as a new alternative method for eye irritation test, protocol refinement was performed and optimum assay condition was determined including treatment time, treatment volume, post-incubation time and rinsing method. Using the refined protocol, 25 reference chemicals with known eye irritation potentials were tested. With the viability cut-off value at 50%, chemicals were classified to irritant or non-irritant. When compared with GHS classification, the MCTT HCE model showed the accuracy of 88%, sensitivity of 100% and specificity of 77%. These results suggest that the MCTT HCE model might be useful as a new alternative eye irritation test method.
Keywords:MCTT HCE model   Eye irritation test   Barrier function   Cell viability assay   Protocol refinement   Predictive model
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