A new 3D reconstituted human corneal epithelium model as an alternative method for the eye irritation test.
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.
Amorepacific Corporation R&D Center, Giheung-gu, Yongin, Republic of Korea.
Kyoung-Mi
KM
Jung
Su-Hyon
SH
Lee
Yang-Hwan
YH
Ryu
Won-Hee
WH
Jang
Haeng-Sun
HS
Jung
Ju-Hee
JH
Han
Seung-Hyeok
SH
Seok
Jae-Hak
JH
Park
Youngsook
Y
Son
Young-Ho
YH
Park
Kyung-Min
KM
Lim
D016475:3T3 Cells; D000826:Animal Testing Alternatives; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D017097:Electric Impedance; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D016850:Limbus Corneae / Q000166:cytology; D051379:Mice; D008954:Models, Biological; D017830:Octoxynol / Q000633:toxicity; D015203:Reproducibility of Results; D013997:Time Factors; D018675:Toxicity Tests / Q000379:methods*
D015415:Biomarkers; D007509:Irritants; D017830:Octoxynol