Experimental Verification of Human Cell Line Activation Test for Skin Allergenicity Detection

Objectives To establish the human cell line activation test (h-CLAT) in laboratory to detect skin allergenicity of dye precursors and hair dye products.

Methods To establish the method and determine the reliability of the experiment by using 10 proficiency substances recommended in ANNEX Ⅱ of OECD 442(E). Four samples were tested for skin allergenicity. The cytotoxicities of the 10 proficiency substances were measured by CCK-8 method and flow cytometry method respectively, and comparisons were further conducted.

Results The result of 10 proficiency substances were in accordance with OECD 442(E) requirements. Four samples' skin allergenicity result of h-CLAT were consistent with those of the LLNA method. The cytotoxicity result of CCK-8 and flow cytometry were both in accordance with the OECD reference range. The intra-group correlation coefficient (ICC) between the two method was 0.993.

Conclusions A reliable h-CLAT test method has been established in the laboratory and can be applied to the allergenicity detection of some hair dye products.