In Vitro Embryotoxicity Test of Four Plasticizers by Embryonic Stem Cell Test (EST)

Koichi Imai1, Shoji Takeda1, Kazuhiko Suese2, Yoshihiro Momota3, Naotaka Kishimoto3, Michiko Nakatsuka4, Shunji Kumabe4, Yasutomo Iwai4, Shinji Kusakawa5, and Akito Tanoue5

1Department of Biomaterials, Osaka Dental University, Osaka, Japan,
2Osaka Dental University. School of Dental Technician and Hygienist, Osaka, Japan,
3Department of Anesthesiology, Osaka Dental University, Osaka, Japan,
4Department of Oral Anatomy, Osaka Dental University, Osaka, Japan,
5Department of Pharmacology, National Research, Institute for Child Health and Development, Tokyo, Japan

AATEX 14(3):947-953, 2009

The embryonic stem cell test (EST) is a well-known in vitro embryotoxicty test protocol. We therefore investigated the embryotoxicity of four plasticizers by the EST protocol. Three parameters (ID50, IC50ES and IC503T3) obtained by the results were substituted for the prediction model for classification. Each embryotoxicity level of DBP, BBP and DEHA was classified into the category of "weak embryotoxicity". Only DEHP was classified as "no embryotoxicity".
As we carried out these experiments, we were able to identify new weak points in the EST protocol; that is, using the high concentration solution of the four plasticizers, it is difficult to make an embryoid body (EB) from the hanging drop culture of the EST protocol. A surface-active agent should be considered when such a phenomenon occurs.
When using special chemical substances, improvement of the EST method from the viewpoint of physical properties is also desired.

Key wards: EST, embryotoxicity, embryoid body, plasticizer


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