A new 3D reconstituted human corneal epithelium model as an alternative method for the eye irritation test

作者：Jung Kyoung Mi; Lee Su Hyon; Ryu Yang Hwan; Jang Won Hee; Jung Haeng Sun; Han Ju Hee; Seok Seung Hyeok; Park Jae Hak; Son Youngsook; Park Young Ho; Lim Kyung Min^*

来源：Toxicology in Vitro, 2011, 25(1): 403-410.

DOI：10.1016/j.tiv.2010.10.019

• MCTT HCE model
• Eye irritation test
• Barrier function
• Cell viability assay
• Protocol refinement
• Predictive model

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 bio-marker 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.

• 出版日期2011-2