Validation Study on Five Cytotoxicity Assays by JSSSE — I. Overview of The Study And Analyses of Variations of ED50 Values
Tadao Ohno1, Masumi Asakura2, Takumi Awago3, Yoshihiro Futamura4, Akihiro Harihara5, Masato Hatao6, Chiharu Hattori7, Akira Hayasaka8, Makoto Hayashi9, Takashi Yahashi10, Zenzou Hirata4, Hiroshi Hori11, Hiroyoshi Hoshi12, Koichi Imai13, Ikuo Imazeki8, Takuya Ishibashi11, Hiroshi Itagaki6, Tatsuo Iwata14, Miyako Kakuma15, Shinya Kaneda16, Ikuo Kato5, Mayako Kato1, Tooru Kawahatsu17, Akio Kawakami18, akemi Kazama19, Akihiko Kido20, Shigemi Kimura21, Michiyo Kitazawa22, Shigefumi Kogiso23, Hajime Kojima24, Mayumi Kotani25, Masahiro Kuramochi21, Daijyo Maki26, Masako Matsuda27, Chihomi Mitsuoka26, Satoru Miyazaki28, Fumio Mizuno20, Minako Mori29, Katsunori Morimoto30, Matsuko Moriysu26, Keiya Nakajima29, Madoka Nakajima22, Masaaki Nakamura13, Masaki Nakamura4, Nahoko Nakano26, Shinobu Nakamura26, Akira Inegami31, Masato Nishino19, Tamotsu Nishitomi20, Kenji Ohkoshi32, Yuuko Okamoto32, Takashi Omori35, Hiroshi Ono34, Makoto Ono15, Yuko Osanai15, Kaoru Saijo1, Yuka Sano35, Kyoko Saotome36, Kazunori Sasaki5, Tetsuji Sasaki1, Hidetaka Sato37, Shinichiro Sato38, Hiroyasu Shimada7, Shoichiro Shimogo6, Kazuyuki Shimono16, Hiroshi Shionoya14, Hideaki Sugawara28, Yasuko Sugiki39, Shigeo Sugimoto40, Shigeru Sugimoto37, Junko Suzuki37, Kinya Takagaki4, Ken Takahashi41, Minoru Takizawa42, Chieko Tamaki23, Noriho Tanaka34, Jirou Taniya34, Noriko Teramoto1, Hisashi Torishima44, Toshiyuki Tsuchiya45, Motoo Uejima16, Hayao Ueno46, Yuji Ugai22, Shogo Wada47, Shinobu Wakuri34, Xinhai Wang1, Ikuo Watanabe29, Masami Watanabe48, Satoshi Yajima49, Yoshiki Yamagata19, Yuuzou Yamaguchi49, Osamu yamakita40, Ryohei Yamamoto44, Masanori Yoshida46, Isao Yoshimura33, Kouichi Yuhki43, Shiro Yukiyama40
1 RIKEN Cell Bank, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukaba Science City, Ibaraki 305;
2 Japan Bioassay Laboratory, 2445 Hirasawa, Hadano-shi, Kanagawa 257;
3 Drug Safety Research Center, Tokushima Research Institute, Otsuka Pharmaceutical Co., Ltd., 463-10 Kagasuno, Kawauchi-cho, Tokushima-shi, 771-01;
4 Toxicology Laboratory, Mochida Pharmaceutical Co., Ltd., 342 Gensuke, Fujieda-shi, Shizuoka 426;
5 Shionogi Research Laboratories, Shionogi & Co., Ltd., 3-1-1 Futaba-cho, Toyonaka-shi, Osaka 561;
6 Shiseido Safety & Analytical Research Center,1050 Nippa-cho, Kohoku-Ku, Yokohama-shi 223;
7 Developmental Research Laboratory, Daiich Pharmaceutical Co., Ltd., 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo 134;
8 Chugai Pharmaceutical Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171;
9 Division of Genetics and Mutagenesis, Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158;
10 Ina Research Inc., 8047 Nishiminowa, Ina-shi, Nagano 399-45;
11 Tsuruga Institute of Biotechnology, Toyobo Co., Ltd., 10-24 Toyo-cho, Tsuruga-shi, Futui 914;
12 Research Institute for the Functional Peptides, 11-26 Minamisanban-cho, Yamagata-shi 990;
13 Department of Biomaterials, Osaka Dental University, 1-5-31 Otemae, Chuo-ku, Osaka-shi 540;
14 Department of Drug Safety Research, R & D Division, Eisai Co., Ltd., Takehaya, Kawashima-cho, Hashima-gun, Gifu 501-61;
15 Beauty Care Products Laboratory, Nagase, Co., Ltd., 5-1 Kobuna-cho, Nihonbashi, Chuo-ku, Tokyo 103;
16 Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc., 115 Kuguhara, Tateiwa, Muya-cho, Naruto-shi, Tokushima 772;
17 Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama-shi 222;
18 Safety Research Laboratories, Yamanouchi Pharmaceutical Co., Ltd., 1-1-8 Azusawa, Itabashi-ku, Tokyo 174;
19 Biochemical Development & Production, Oriental Yeast Co., Ltd., 4-4-1 Minamisuita, Suita-shi, Osaka 564;
20 Kashima Laboratory, Mitsubishi-kagaku Institute of Environmental and Toxicological Sciences, 14 Sunayama, Hasaki-machi, Kashima-gun, Ibaraki 314-02;
21 BML Inc., 1361-1 Matoba, Kawagoe-shi, Saitama 350;
22 Biosafety Research Center, Foods, Drugs and Pesticides, 582-2 Arahama, Shioshinden, Fukude-cho, lwata-gun, Shizuaka 437-12;
23 Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 3-1-98 Kasugadanaka, Konohana-ku, Osaka-shi 554;
24 Biochemical Research Institute, Nippon Menard Cosmetic Co., Ltd., 2-7 Torimi-cho, Nishi-ku, Nagoya-shi 451;
25 R&D Planning HQ, Sunstar Inc., 3-1 Asahi-machi, Takatsuki-shi, Osaka 569;
26 Panapharm Laboratories Co., Ltd. 1285 Kurisaki-cho, Uto-shi, Kumamoto 869-04;
27 Research and Development Center, Duskin Co., Ltd., 4-19-8 Minamisuita, Suita-shi, Osaka 564;
28 Life Science Research Information Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-01;
29 The Center of Japan Biological Chemistry, 52 Fukue, Kaizu-cho, Kaizu-gun, Gifu 503-06;
30 Takara Belmont Co., Ltd., 5-1 Takamatsu, Kouzai-cho, Koga-gun, Shiga 520-32;
31 Deparment of Obstetrics and Gynecology, Fukui Medical School, 23-3 Shimoaizuki, Matsuoka-cho, Yoshida-gun, Futui 910-11;
32 Division of Fundamental Research, KOSE Co., 1-18-4 Azusawa, Itabashi-ku, Tokyo 174;
33 Department of Management Science, Faculty of Engineering, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162;
34 Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257;
35 Biotechnology Research Laboratories, TAKARA SHUZO Co., Ltd., 3-4-1 Seta, Ohtsu-shi, Shiga 520-21;
36 Yokohama City Institute of Health, 1-2-17 Takigashira, Isogo-ku, Yokohama-shi 235;
37 Japan Food Research Laboratories, 6-11-1O Nagayama, Tama-shi, Tokyo 206;
38 Division of Radiation Medicine, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba-shi 263;
39 Menicon Co., Ltd., 5-1-10 Takamoridai, Kasugai-shi, Aichi 487;
40 Drug Safety Laboratory, Taiho Pharmaceutical Co., Ltd., 224-2 Ebisuno, Hiraishi, Kawauchi-Cho, Tokushima-shi 771-01;
41 The Institute of Environmental Toxicology, 4321 Uchimoriya-cho, Mitsukaido-shi, Ibaraki 303;
42 Hakko Electric Machine Works Co. Ltd., 3055 Togura, Toguramachi, Hanishina-gun, Nagano 389-08;
43 Koken Bioscience Institute, Koken Co., Ltd., 2-11-12 Nakane, Meguro-ku, Tokyo 152;
44 Kurabo Industries Ltd., 14-41 Shimokida-cho, Neyagawa-shi, Osaka 572;
45 Safety Evaluation Center, Central Research Laboratory, Showa Denko K K, 1-1-1 Ohnodai, Midori-ku, Chiba-shi 267;
46 Takeda Analytical Research Laboratories, Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka 532;
47 Wako Pure Chemical Industries, Ltd., 6-1 Takadacho, Amagasaki-shi, Hyogo 661;
48 Division of Radiation Biology, Department of Health Science, Faculty of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki-shi 852;
49 Central Research Laboratory, Takasago International Co., 1-4-11 Nishiyawata, Hiratsuka-shi, Kanagawa 254, Japan
(pp. 1-38)
Validation Study on Five Cytotoxicity Assays by JSAAE — II. Statistical Analysis
Takashi Omori1, Kaoru Saijo2, Mayako Kato2, Hiroshi Itagaki3, Makoto Hayashi4, Satoru Miyazaki5, Tadao Ohno2, Hideaki Sugawara5, Noriko Teramoto2, Noriho Tanaka6, Shinobu Wakuri6 and Isao Yoshimura1
1 Department of Management Science, Faculty of Engineering, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162;
2 RIKEN Cell Bank, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukaba Science City, Ibaraki 305;
3 Shiseido Safety & Analytical Research Center,1050 Nippa-cho, Kohoku-Ku, Yokohama-shi 223; 4 Division of Genetics and Mutagenesis, Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158;
5 Life Science Research Information Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-01;
6 Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257
(pp. 39-58)
Validation Study on Five Cytotoxicity Assays by JSAAE — III. Quality of Collected Data Files
Takashi Omori1, Kaoru Saijo2, Mayako Kato2, Hiroshi Itagaki3, Makoto Hayashi4, Satoru Miyazaki5, Tadao Ohno2, Hideaki Sugawara5, Noriko Teramoto2, Noriho Tanaka6, Shinobu Wakuri6, and Isao Yoshimura1
1Department of Management Science, Faculty of Engineering, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162;
2RIKEN Cell Bank, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyodai Tsukuba Science City, Ibaraki 305;
3Shiseido Safety & Analytical Research Center, 1050 Nippa-cho, Kohoku-ku, Yokohama-shi 223;
4Division of Genetics and Mutagenesis, Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158;
5Life Science Research Information Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-01;
6Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257, Japan
(pp. 59-73)
Validation Study on Five Cytotoxicity Assays by JSAAE — IV. Details of The Colony Formation Assay
Noriho Tanaka1, Masumi Asakura2, Chiharu Hattori3, Akira Hayasaka4, Makoto Hayashi5, Takashi Yahashi6, Hiroshi Hori7, Hiroyoshi Hoshi8, Ikuo Imazeki4, Takuya Ishibashi7, Hiroshi Itagaki9, Miyako Kakuma10, Shinya Kaneda11, Mayako Kato1, Akio Kawakami12, Akihiko Kido13, Michiyo Kitazawa14, Hajime Kojima15, Daijyo Maki16, Chihomi Mitsuoka16, Satoru Miyazaki17, Fumio Mizuno13, Matsuko Moriysu16, Madoka Nakajima14, Nahoko Nakano16, Shinobu Nakamura16, Tamotsu Nishitomi14, Tadao Ohno18, Masato Omori19, Hiroshi Ono1, Makoto Ono10, Yuko Osanai10, Kaoru Saijo18, Tetsuji Sasaki18, Hidetaka Sato20, Shinichiro Sato21, Hiroyasu Shimada3, Kazuyuki Shimono11, Hideaki Sugawara17, Yasuko Sugiki22, Shigeo Sugimoto20, Shigeru Sugimoto20, Junko Suzuki20, Ken Takahashi23, Minoru Takizawa24, Jirou Taniya24, Noriko Teramoto18, Toshiyuki Tsuchiya26, Motoo Uejima11, Hayao Ueno27, Yuji Ugai14, Shinobu Wakuri1, Xinahi Wang18, Masanori Yoshida27, Isao Yoshimura19, Kouichi Yuhki25
1Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257;
2Japan Bioassay Laboratory, 2445 Hirasawa, Hadano-shi, Kanagawa 257;
3Developmental Research Laboratory, Daiich Pharmaceutical Co., Ltd., 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo 134;
4Chugai Pharmaceutical Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171;
5Division of Genetics and Mutagenesis, Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158;
6Ina Research Inc., 8047 Nishiminowa, Ina-shi, Nagano 399-45;
7Tsuruga Institute of Biotechnology, Toyobo Co., Ltd., 10-24 Toyo-cho, Tsuruga-shi, Futui 914;
8Research Institute for the Functional Peptides, 11-26 Minamisanban-cho, Yamagata-shi 990;
9Shiseido Safety & Analytical Research Center,1050 Nippa-cho, Kohoku-Ku, Yokohama-shi 223;
10Beauty Care Products Laboratory, Nagase, Co., Ltd., 5-1 Kobuna-cho, Nihonbashi, Chuo-ku, Tokyo 103;
11Naruto Research Institute, Otsuka Pharmaceutical Factory, Inc., 115 Kuguhara, Tateiwa, Muya-cho, Naruto-shi, Tokushima 772;
12Safety Research Laboratories, Yamanouchi Pharmaceutical Co., Ltd., 1-1-8 Azusawa, Itabashi-ku, Tokyo 174;
13Kashima Laboratory, Mitsubishi-kagaku Institute of Environmental and Toxicological Sciences, 14 Sunayama, Hasaki-machi, Kashima-gun, Ibaraki 314-02;
14Biosafety Research Center, Foods, Drugs and Pesticides, 582-2 Arahama, Shioshinden, Fukude-cho, lwata-gun, Shizuaka 437-12;
15Biochemical Research Institute, Nippon Menard Cosmetic Co., Ltd., 2-7 Torimi-cho, Nishi-ku, Nagoya-shi 451;
16Panapharm Laboratories Co., Ltd. 1285 Kurisaki-cho, Uto-shi, Kumamoto 869-04;
17Life Science Research Information Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-01;
18RIKEN Cell Bank, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukaba Science City, Ibaraki 305;
19Department of Management Science, Faculty of Engineering, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162;
20Japan Food Research Laboratories, 6-11-1O Nagayama, Tama-shi, Tokyo 206;
21Division of Radiation Medicine, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba-shi 263;
22Menicon Co., Ltd., 5-1-10 Takamoridai, Kasugai-shi, Aichi 487;
23The Institute of Environmental Toxicology, 4321 Uchimoriya-cho, Mitsukaido-shi, Ibaraki 303;
24Hakko Electric Machine Works Co. Ltd., 3055 Togura, Toguramachi, Hanishina-gun, Nagano 389-08;
25Koken Bioscience Institute, Koken Co., Ltd., 2-11-12 Nakane, Meguro-ku, Tokyo 152;
26Safety Evaluation Center, Central Research Laboratory, Showa Denko K K, 1-1-1 Ohnodai, Midori-ku, Chiba-shi 267;
27Takeda Analytical Research Laboratories, Ltd., 2-17-85 Juso-honmachi, Yodogawa-ku, Osaka 532
Original paper :AATEX 5(1-2):74-86,1998
Abstract
The inter-laboratory validation study on 5 cytotoxicity assays conducted by JSAAE has been described in the preceding articles in this issue. Presented here are precise data and the protocols on the colony formation (CF) assay with two cell lines, HeLa S3 (SC) that is common to the other 4 assays and BALB/3T3 A31-1-1 that is frequently used in this assay. The CF assay showed high performance rates with both HeLa S3 (SC) and BALB/3T3 A31-1-1 cell lines. Ninety-five percent of submitted data files were acceptable before ED50 calculation in this assay. Variations on negative controls in the assay revealed technical differences among laboratories and the characteristics of cells applied. The CF assay with BALB/3T3 A31-1-1 cells resulted in the lowest median values of log(ED50) for Tween 20, Tween 80 and cetylpyrimidinium chloride monohydrate. For propylene glycol assayed with HeLa S3 (SC) cells, the lowest median value of log(ED50) was observed. Thus the CF assay was able to detect with the highest sensitivity the least toxic chemical (propylene glycol) and the most severely toxic chemical (cetylpyrimidinium chloride monohydrate). CF assay with HeLa S3 (SC) cells gave the largest power for distinction when observing the most and the least toxic chemicals. However, the toxicities of moderately toxic chemicals were not sharply distinguishable from each other. Comparing the mean hinge-spreads, CF assay with BALB/3T3 A31-1-1 cells showed the largest variation of the log(ED50) values among all assay systems examined in this study. These results suggest that the CF assay is a highly sensitive assay but is influenced by many factors such as culture conditions and techniques of cell handling.
Key words: alternatives, colony formation assay, cytotoxicity assay, inter-laboratory validation