Embryotoxicity of the Multi-walled Carbon Nanotubes (MWCNTs) using the Three-dimensional Culture of ES-D3 cells

44 Introduction An increasing number of nano or submicron materials have been developed . However, their effects on human health remain unclear. Thus, the development of a safety evaluation method is an urgent issue . No embryotoxicity test on teratogenicity in humans has been developed. We have investigated the applications of the EST (Embryonic Stem Cell Test) protocol, i.e., an in vitro embryotoxicity test whose predictability has been established by international validation in the U.S. and Europe , to nano or submicron materials. We have already reported the effects of the multi-walled carbon nanotubes (MWCNTs) on the differentiation of ES-D3 cells 16, . However, it was intended to differentiate ES-D3 cells on a dish with MWCNTs adsorbed on the bottom, but not to differentiate cells from embryonic bodies as described in the EST protocol. In the present study, we compared three-dimensional culture using a collagen gel with conventional two-dimensional culture. Embryotoxicity of the Multi-walled Carbon Nanotubes (MWCNTs) using the Three-dimensional Culture of ES-D3 cells