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

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

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

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.

Original languageEnglish
Pages (from-to)403-410
Number of pages8
JournalToxicology in Vitro
Volume25
Issue number1
DOIs
StatePublished - Feb 2011

Keywords

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

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