Development of alternatives to animal experiments using pluripotent stem cells

Abstract

Animal experiments have occupied an important position in the safety assessment of chemicals. However, due to the rise in animal welfare as seen in the ban of animal experiments in European cosmetic development, the development of alternative methods for animal experiments has become very important in recent years. Development of in vitro tests for local toxicity such as irritation and sensitization tests is preceded. Meanwhile, alternative tests for systemic toxicity such as chronic and developmental toxicities are under development. In developing alternative methods using cultured cells, we have been focusing on pluripotent stem cells such as ES and iPS cells and studying alternatives to developmental toxicity and neurotoxicity. As an alternative test of developmental toxicity, we developed the Hand 1-Luc EST, which is a simple test utilizing cardiomyocyte differentiation process of mouse ES cells, and Tubb 3- and Reln-Luc ESTs using nerve differentiation process. Recently, it was clarified that the combination of the Hand 1-Luc EST and the Tubb 3- and Reln-Luc ESTs improves the prediction of the developmental toxicity. In the study of in vitro neurotoxicity test using neurons derived from mouse ES cells, evaluation methods for neurite outgrowth using high-content imaging technology and for neural function using multi-electrode arrays were developed. In addition, we introduce differentiation methods for retinal tissues from human ES/iPS cells, which are the results as the collaboration with RIKEN and the present state of an in vitro phototoxicity test using retinal pigment epithelial cells (RPE) derived from human ES cells.