Proceedings from the 8th Congress of KSAAE

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Proceedings from the 8th Congress of KSAAE

Published: August 1, 2011
The Eighth Congress of the Korean Society for Alternatives to Animal Experiments (KSAAE) was held on July 8, 2011 at Hoseo University, South Korea. Three morning presentations described research and developments of alternative test methods in Korea, Japan, and China. The abstracts of the three presentations from the morning session are reprinted in full below.

Current and future collaborations between Japan and Korea on alternatives to animal experiments

By Hajime Kojima, Japanese Center for the Validation of Alternative Methods, National Institute of Health Sciences, Tokyo, Japan


The Japanese Society for Alternatives to Animal Experiments (JSAAE)1 and the Korean Society for Alternatives to Animal Experiments (KSAAE)2 have had a good relationship prior to and after the 6th World Congress on Alternatives and Animal Use in the Life Sciences (WC6) in August, 2007. The satellite symposium of WC6 was held in Seoul and JSAAE invited a large number of Korean scientists. Furthermore, the Korean Center for the Validation of Alternative Methods (KoCVAM)3, which was established by the Korean Food and Drug Association (KFDA) in November, 2009, has initiated international collaborations with validation centers around the world.

The establishment of alternatives to animal testing necessitates international collaborations. To this point, international organizations including the Organization for Economic Co-operation and Development (OECD), International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH), and International Cooperation on Cosmetics Regulations (ICCR), have expressed a need for international cooperation in order to develop, validate, and review new test guidelines and regulations. Currently, international test guidelines are not a national rule in Japan and Korean, although they should be followed worldwide. In order to achieve this aim, the International Cooperation on Alternative Test Methods (ICATM)4 has been organized among the EU, USA, Canada, Japan, and Korea. ICATM was established to expand and strengthen cooperation, collaboration, and communications among national validation organizations to further the scientific validation and evaluation of new alternative testing methods proposed for regulatory health and safety assessments.


To date, a limited number of collaborations between Japan and Korean researchers have been conducted in this field. For example, the Japanese Center for the Validation of Alternative Methods (JaCVAM)5 is collaborating with Korean colleagues on validation studies of ERα STTA antagonist assay using HeLa9903 and in vitro comet assay. Although the aforementioned collaboration between JaCVAM and Korean colleagues has been successful, these are insufficient and further cooperation among Asian countries should be established. Moreover, a mutual relationship between Japan and Korea should be encouraged in order to develop new alternatives to animal methods, as these two countries are at the intersection of Asia and the rest of the world.

New multicolor reporter gene assay for toxicity test

By Yoshihiro Ohmiya1, Shigeaki Nishii2, Tomomi Yarnazaki3, Yoshihiro Nakajiam2

1National Institute of AIST, Bioproduction Research Institute, Tsukuba, Japan
2National Institute of AIST, Health Research Institute, Takamatsu, Japan
3TOYOBO CO., LTD., Tsuruga, Japan


In the post genome era, reporter assay systems are used widely to study promoters, interactions between promoters and transcription factors, signal transduction and other cellular activities. Reporter assays are also applied to drug and toxicity screening in vitro, in cellulo, and in vivo. Of the reporter genes known to date, luciferases, enzymes that catalyze bioluminescence reactions, are used most frequently because their sensitivity and linear response range are superior to those of typical reporters including ß-galactosidase, chloramphenicol acetyltransferase, ß-glucuronidase and fluorescent proteins. Bioluminescence is a simple reaction that is triggered by the addition of luciferin solution, and the equipment for measuring light intensity is simple because it uses only a photomultiplier or a charge-coupled device (CCD) camera. So, luciferases are suitable reporter enzymes for the quantitative measurement of gene expression. In this section, I introduce our creating system using multiple bioluminescence probes. The multicolor reporter assay using beetle luciferases that emit various colors with a luciferin can observe the dynamics of three gene expressions in the cells. Namely, our tricolor reporter in vitro assay system in which three gene expressions are monitored simultaneously using green-, orange- and red-emitting beetle luciferases for gene expression analysis. The multicolor reporter cell line reveals the expressions change of two or three genes under the target chemicals simultaneously. Our technique is a unique and a powerful tool for the toxicity test.

China’s Alternative Test Methods Research in Cosmetics: Current Situation and Prospects

By Xikun Xiong, Vice Director of Toxicology Institute Guangdong CDC, Guangzhou, China

This article describes the Chinese history of alternative methods of animal testing, introduces China’s current situation in research on alternative methods, and forecasts the application of alternative methods of animal testing in China.

In 1997, in China development plan of experimental animals, the former China state science and technology commission, for the first time, wrote “3 Rs” basic concept into Experimental animal management and technology development files. Since then, six institutions including university, research institution, CDC, and so on successively developed alternative test animal research work. At present, alternative animal testing is mainly implicated in cosmetics and ingredient safety evaluation. In 2008, Guangdong CDC undertook the research project funded by the health ministry special funds “Alternative Animal Testing and Toxicology Safety Evaluation and Research”, it has made positive progress in leading Chinese alternative animal testing’s establishment and application.

At present, China is not against alternatives to animal testing, but China has no clear laws and regulations to approve alternative animal testing. The author thinks that, in the near future, China may constitute some Acts or national standards for animal alternative tests, but China will not ban animal testing.

1 JSAAE (2011) Available at:
2 KSAAE (2001) Available at:
3 KoCVAM (2011) Available at:
4 ICATM (2011) Available at:
5 JaCVAM (2011) Available at:

Nakajima, Y., Ikeda, M., Kimura, T., Honma, S., Ohmiya, Y. & Honma, K. (2004). Role of orphan nuclear receptor ROR a in clock gene transcriptions demonstrated by a novel reporter assay system. FEBS Lett. 565, 122-126.

Nakajima, Y., Kimura, T., Sugata, K., Enomoto, T., Asakawa, T., Kubota, H., Ikeda, M. & Ohmiya, Y. (2005). A multicolor luciferase assay system, one-step monitoring of multiple gene expressions with a single substrate. Biotechniques. 38, 891-894.

Li, X., Nakajima, Y., Niwa, K., Viviani, V.R. & Ohmiya, Y. (2010). Enhanced red-emitting railroad worm luciferase for bioassays and bioimaging. Protein Sci. 19, 26-33.

Nakajima, Y., Yamazaki, T., Nishii, S., Noguchi, T., Hoshino, H., Niwa, K., Viviani, V.R. & Ohmiya, Y. 2010. Enhanced beetle luciferase for high-resolution bioluminescence imaging. PLoS One. Apr 2;5(4):e10011.

Kwon, H.J., Enomoto, T., Shimogawara, M., Yasuda, K., Nakajima, Y. & Ohmiya, Y. (2010). Bioluminescence imaging of dual gene expression at the single-cell level. BioTechniques. 48(6), 460-2.