In vitro eye irritation test (OECD TG 492)
Reconstructed human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eyedamage.
Eye irritation test – Artificial cornea
Using the human corneal-like epithelium (RhCE) model, which closely mimics the histological, morphological, biochemical, and physiological characteristics of the human corneal epithelium, the assessment of eye irritation (UN GHS Category 2) or severe eye damage (UN GHS Category 1) is conducted according to UN GHS guidelines.
This evaluation is utilized to assess the toxicity of testsubstances (compounds and mixtures) that do not require classification andlabeling for UN GHS Category 1 (UN GHS No Category).
Test Process
Step 01
Preculture
Medium 0.9ml 37°C, 5% CO2, 22±2hours
Step 02
Test sample treatment
Liquide: 40μl. 10min Solid: DPBS 40μl + 40mg, 37°C, 5% CO2, 3hours
Step 03
Wash
Washing the inside and outside of the human cornea model with DPBS
Step 04
Post-culture
Medium 0.9ml 37°C, 5% CO2, 16±1hour
Step 05
WST-1 assay
WST-1 solution 300μl (insert inside 100μl, insert outside 200μl) 37°C, 5% CO2, 3hours±5min
Step 06
Absorbance Measurement
Supernant 200μl / well(96 well plate) 450nm