ORIGINAL ARTICLE
A Study of the Criteria for Selection of THP-1 Cells in the Human Cell Line Activation Test (h-CLAT): Results of 2nd Japanese Inter-laboratory Study

Nanae Kosaka1, Kenji Okamoto2, Makoto Mizuno3, Takaaki Yamada4, Mayumi Yoshida5, Tatsuji Kodama6, Sono Sakiko7, Takao Ashikaga7, Jun Sato6, Naoko Ota5, Seiji Hasegawa4, Yuko Okamoto3, Hirofumi Kuwahara2, Hitoshi Sakaguchi1 and Yasuo Ohno8

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

AATEX 13(2):55-62, 2008

Abstract
The human Cell Line Activation Test (h-CLAT) is an in vitro skin sensitization method based on augmentation of CD86 and CD54 expression in THP-1 cells (human monocytic leukemia cell line). In our previous Japanese inter-laboratory study, we reported that the transferability and reproducibility of the h-CLATis basically good. The aim of this study was to define the criteria for selecting appropriate THP-1 cells inthe h-CLAT. In this study, new THP-1 cell lots were obtained from three cell banks: one in America,Europe and Japan. Using these three lots plus the cell lot we had previously used and obtained from ATCCwe investigated the CD86/CD54 expression following exposure to two allergens (DNCB and Ni) and onenon-allergen (SLS). Compared with the previous ATCC lot, two new lots showed similar results. Mean-while, the third new lot showed distinctly different results in cell viability and CD86/CD54 augmentationinduced by Ni compared to the other three lots. These results showed that the variability of cellular responses in the THP-1 cells depended on the cell source. In conducting the h-CLAT, it would be importantto select appropriate THP-1 cells to predict correctly the skin sensitization potential.

Key words: h-CLAT, skin sensitization, alternatives, THP-1, cell selection

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