ORIGINAL ARTICLE
A study on serum difference on test results in the human Cell Line Activation Test (h-CLAT): Results of 3rd Japanese inter-laboratory study

Sakiko Sono1, Takaaki Yamada2, Nanae Kosaka3, Kenji Okamoto4, Makoto Mizuno5, Jun Sato6, Mayumi Yoshida7, Naoko Ota7, Tatsuji Kodama6, Yuko Okamoto5, Hirofumi Kuwahara4, Hitoshi Sakaguchi3, Seiji Hasegawa2, Takao Ashikaga1 and Yasuo Ohno8

1Shiseido Co., Ltd., 2Nippon Menard Cosmetic Co., Ltd., 3Kao Corporation, 4Kanebo Cosmetics Inc., 5KOSÉ Corporation, 6LION CORPORATION, 7POLA CHEMICAL INDUSTORIES, INC., 8National Institute of Health Sciences, Japan

AATEX 13(2):63-69, 2008

Abstract
The human Cell Line Activation Test (h-CLAT) is an in vitro skin sensitization test based on the enhancement of CD86 and/or CD54 expression on THP-1 cells. The aim of this study was to examine the effect of differences of serum source on the results of h-CLAT. Three different lots of serum, obtained from three sources, were compared with the serum used in the previous Japanese ring study. With each serum, cellular proliferation in subculture, cytotoxicity, and CD86/CD54 expression on THP-1 cells were measured following exposure to two known allergens (dinitrochlorobenzene (DNCB) and nickel sulfate (Ni)) and one non-allergen (sodium lauryl sulfate (SLS)).
There was no clear difference of cellular proliferation in subculture, cytotoxicity, or CD86/54 expression among cultures in the four sera. Although the source of serum does not appear to influence the resultof h-CLAT, the validity of the test should nevertheless be confirmed when serum from a new source is introduced.

Key words: skin sensitization, alternative test, serum, h-CLAT

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(AATEX: Altern. Animal Test. EXperiment.: Alternatives to Animal Testing and EXperimentation)