ORIGINAL MANUSCRIPT
In Vitro Embryotoxicity Testing of Mercury Vapour by Differentiation of ES-D3 Cells

Koichi Imai1, Shinji Kusakawa2, Akito Tanoue2, Makiko Kuwagata3, Mika Senuma3, Mami Furuya3 and Hiromasa Takashima3

1Department of Biomaterials, Osaka Dental University, Osaka, Japan, 2Department of Pharmacology, National Research Institute for Child Health and Development, Tokyo, Japan, 3Laboratory of Toxicology, Hatano Research. Institute., FDSC, Kanagawa, Japan

AATEX 13(3):118-122, 2008

Abstract
The embryonic stem cell test (EST) is an in vitro assay that has been developed to assess the embryotoxic potential of chemicals and biomaterials. It is based on the capacity of the mouse embryonic stem cell line D3 (ES-D3 cells) to differentiate into contracting myocardial cells. However, the EST method cannot fully test the embryotoxicity of mercury vapor. We tried to apply the rotary culture bottle containing mercury vapor and culture medium was slowly turned and the cells come into direct contact with the gaseous phase containing mercury vapor. At the mercury exposure concentrations tested in this study, no cell pulsations were observed. The cell viability was affected to some extent in the 5.0µg mercury concentration group, whereas in the 2.0µg group little difference was observed from that of the control group. Although mercury vapor was observed to strongly impair the differentiation of EBs in this study, the test results this time did not prove that mercury vapor has a strong embryotoxic effect on humans. We think it important to develop a device that closely mimics the actual exposure conditions of the human body, including an exposure method that is different from that used in this study, as an alternative to animal experiments.

Key words: mercury vapour, embryotoxicity, ES cell

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