Miniaturized and multiplexed high-content screening of drug and immune sensitivity in a multichambered microwell chip

Here, we present a methodology based on multiplexed fluorescence screening of two-or three-dimensional cell cultures in a newly designed multichambered microwell chip, allowing direct assessment of drug or im-mune cell cytotoxic efficacy. We establish a framework for cell culture, formation of tumor spheroids, fluores-cence labeling, and imaging of fixed or live cells at various magnifications directly in the chip together with data analysis and interpretation. The methodology is demonstrated by drug cytotoxicity screening using ovarian and non-small cell lung cancer cells and by cellular cytotoxicity screening targeting tumor spheroids of renal carcinoma and ovarian carcinoma with natural killer cells from healthy donors. The miniaturized format allowing long-term cell culture, efficient screening, and high-quality imaging of small sample volumes makes this methodology promising for individualized cytotoxicity tests for precision medicine.

Automated summary

In this study, a methodology based on a multichambered microwell chip is presented, which allows for direct assessment of drug or immune cell cytotoxic efficacy. The methodology enables long-term cell culture, efficient screening, and high-quality imaging. The feasibility of this methodology is demonstrated through drug cytotoxicity screening and cellular cytotoxicity screening experiments.