Background Since 3D cell culture models of cancer are a promising way to bridge the gap by better mimicking the in vivo conditions of different organ systems and offering a more accurate representation of physiological responses to drugs and toxins, they have grown exponentially in the past decade, with a broad application in the field of drug discovery. However, due to the complexity of 3D cell culture, there are still a lot of challenges including standardization of techniques and analyzing them, especially saving the cost and time by high throughput screens. Therefore, it is unmet for research to be done to develop a high throughput screening platform that can accurately represent in vivo microenvironment and disease pathology. In this study, we validated a 3D tumor spheroid model platform as a high throughput screening tool to evaluate antibody drug conjugates (ADCs) in cancers.

Methods Generation and optimization of the culture conditions tumor spheroids were performed by using commercially available 3D technology and Ultra low^TM 384-well plates from established cancer cell lines. The responses of tumor spheroids to treatment of antibody ADCs were evaluated by cell-titer glow (CTG) bioluminescence assay.

Results Using Corning low attachment plate, all tested cell lines such as PA-1, Miapaca-2 and A549 cell lines formed a very good tumor spheroid whose size depends on the number of cells seeding number. The CTG assay works on these tumor spheroids. PA-1 and Miapaca-tumor spheroids response ADCs treatment beyond its control. But A549 tumor spheroids didn’t separate the tested ADCs vs its control.

Conclusions We validated a 3D cell culture high throughput platform for ADCs drug screens. This holds promise for future testing of treatment responses, overcoming some of the ethical and scientific disadvantages of using animal models in cancer research.