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 immune cell cytotoxic efficacy. We establish a framework for cell culture, formation of tumor spheroids, fluorescence 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. [Display omitted] •A multichambered microwell chip is presented for miniaturized live cell assays•Assays combine long-term, multiplexed screening with high-resolution imaging•Drug and immune cell cytotoxicity are evaluated in 2D and 3D cancer cell cultures•The method is aimed at precision medicine, cell therapy, or drug development Personalized testing of drugs or immunotherapy is vital for the implementation of precision medicine. Although the field has developed tremendously in recent years, there is still a lack of methods that take advantage of benefits, such as low consumption of patient material or reagents, offered by miniaturized in vitro analysis systems. In this work, Sandström et al. have developed a miniaturized method for high-content screening of drug and immune cell sensitivity of cancer cells. It is advantageous for accurate, time-resolved, and high-resolution analysis of dynamic cellular events and is well suited for applications within personalized medicine.