Validation Study on Five Cytotoxicity Assays by JSAAE -- VI. Details of LDH Release Assay

Tadao Ohno1, Yoshihiro Futamura2, Akihiro Harihara3, Masato Hatao4, Makoto Hayashi5, Zenzou Hirata2, Koichi Imai6, Hiroshi Itagaki3, Mayako Kato1, Tooru Kawahatsu9, Shigefumi Kogiso10, Satoru Miyazaki11, Masaaki Nakamura6, Masaki Nakamura2, Takashi Omori12, Hiroshi Ono13, Makoto Ono8, Yuko Osanai8, Kaoru Saijo1, Yuka Sano14, Kazunori Sasaki3, Tetsuji Sasaki1, Shinichiro Sato15, Shoichiro Shimogo4, Hiroshi Shionoya7, Hideaki Sugawara11, Kinya Takagaki2, Chieko Tamaki10, Noriho Tanaka13, Noriko Teramoto1, Shogo Wada16, Shinobu Wakuri13, Xinhai Wang1, Isao Yoshimura12

1RIKEN Cell Bank, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukaba Science City, Ibaraki 305;
2Toxicology Laboratory, Mochida Pharmaceutical Co., Ltd., 342 Gensuke, Fujieda-shi, Shizuoka 426;
3Shionogi Research Laboratories, Shionogi & Co., Ltd., 3-1-1 Futaba-cho, Toyonaka-shi, Osaka 561;
4Shiseido Safety & Analytical Research Center,1050 Nippa-cho, Kohoku-Ku, Yokohama-shi 223;
5Division of Genetics and Mutagenesis, Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158;
6Department of Biomaterials, Osaka Dental University, 1-5-31 Otemae, Chuo-ku, Osaka-shi 540;
7Department of Drug Safety Research, R & D Division, Eisai Co., Ltd., Takehaya, Kawashima-cho, Hashima-gun, Gifu 501-61;
8Beauty Care Products Laboratory, Nagase, Co., Ltd., 5-1 Kobuna-cho, Nihonbashi, Chuo-ku, Tokyo 103;
9Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama-shi 222;
10Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 3-1-98 Kasugadanaka, Konohana-ku, Osaka-shi 554;
11Life Science Research Information Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-01;
12Department of Management Science, Faculty of Engineering, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162;
13Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257;
14Biotechnology Research Laboratories, TAKARA SHUZO Co., Ltd., 3-4-1 Seta, Ohtsu-shi, Shiga 520-21;
15Division of Radiation Medicine, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba-shi 263;
16Wako Pure Chemical Industries, Ltd., 6-1 Takadacho, Amagasaki-shi, Hyogo 661

Original paper :AATEX 5(1-2):99-118,1998

Abstract
The inter-laboratory validation study on 5 cytotoxicity assays conducted by JSAAE has been described in the preceding articles (see Validation Articles I-V). Presented here are precise data and the protocol of the LDH release assay with two cell lines, HeLa S3 (SC) that is common to the other 4 assays and SQ-5 that is rich in LDH activity. The performance rates in each of the four sub-divided LDH release assays were extremely low. We concluded that the present assay is practically immature, although the assay is very informative in principle not only on the cell growth inhibition but also on the cell Iysis caused by the tested chemicals.

Key words: Key words: alternatives, cytotoxicity assay, inter-laboratory validation, JSAAE project, MTT assay.


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