Effects of Experimental Silver Alloy Containing Indium on the Differentiation of Mouse ES Cells using Artificial Saliva

• Published in: Alternatives to Animal Testing and Experimentation 2019, Vol.24(2), pp.67-74
• Main Authors: Imai, Koichi, Kashiwagi, Kousuke, Han, Xiaoyu, Shirai, Tsubasa, Hashimoto, Yoshiya
• Format: Article
• Language: eng ; jpn
• Subjects: Ag-In alloy; artificial saliva; differentiation; ES cell; indium
• ISSN: 1344-0411; 2185-4726
• DOI: 10.11232/aatex.24.67

The rate of ES-D3 cell differentiation into cardiomyocytes is an important factor in the embryo stem cell tests (EST) protocol for the screening of embryotoxicity. We investigated the effects of dental Ag-In alloys on ES-D3 cells by adding artificial saliva.In the present study, we used Greenwood-prescribed artificial saliva and clinical artificial saliva (Saliveht®) to examine the effect of saliva on the differentiation of ES-D3 cells that are commonly used in EST protocol.We demonstrated that the two artificial types of saliva affected the differentiation of ES-D3 cells when added to the culture medium at 10% of the total volume. We examined the effect of indium on ES-D3 cell differentiation in a collagen-based 3D cell culture system. To do this, we prepared Ag-In alloys with 10, 15, and 20% indium as well as its powdered acid corrosion product and added them to a culture medium that contained artificial saliva at 10% of the total volume. We demonstrated that while cells differentiated at a relatively low level when cultured with an Ag-In alloy that had not been exposed to acid, cells did not differentiate when cultured with the acid corrosion product of the alloy. These findings suggest that the composition of a culture medium has a significant effect on cell differentiation.