Development of a microfluidic device and organoid-based system for evaluation of anticancer drug sensitivity and side effects

Abstract

In this study, for the purpose of evaluate of anti-cancer drug sensitivity and side effects in an environment closer to the living body, mammary tumor organoids produced from mammary tumor affected cats and normal intestinal organoids from mice were perfused with the anticancer drug toceranib after being mounted on a microfluidic device, and comparisons with the non-perfusion condition were made. Analysis was performed.　Cell viability was analyzed after perfusion of cat mammary tumor organoids with culture medium treated with 10 µM toceranib at 9.5 µl / min for 48 hours, and survival was significantly reduced in organoids perfused with the anticancer drug. In addition, the expression of apoptosis-related genes such as p53 and caspase9 was elevated in mammary tumor organoids of cats perfused with toceranib. Next, mammary tumor organoids and mouse normal intestinal organoids were mounted on the same device and perfused with toceranib under the same conditions. Similar to mammary tumor organoids, the survival rate of normal intestinal organoids was also decreased by perfusion of toceranib. However, apoptosis-related genes were not upregulated; instead, the expression of necrosis-related genes such as RIPK3 was increased. Furthermore, intestinal organoids perfused simultaneously with the necrosis inhibitor HS-1371 and toceranib showed a trend toward inhibition of necrosis and increased survival. These results suggest that perfusion with toceranib induces apoptosis in mammary tumor cells and necrosis in normal intestinal cells. They also revealed the usefulness of organoid-loaded microfluidic devices.