Protocol for validating drug efficacy and safety in scalable 96-well platforms of human cortical organoids and melanoma brain metastases.

Metastatic colonization at secondary organs, such as the brain, represents a critical therapeutic window in cancer progression. Here, we present a protocol for a scalable in vitro screening platform for melanoma brain metastases. We describe steps for generating human cortical organoids in 96-well plates, followed by co-culture with melanoma cells to mimic the colonization process. This platform enables quantitative assessment of drug-induced metastasis regression and safety testing using fluorescence and luminescence readouts, targeting colonized melanoma cells in the brain microenvironment. For complete details on the use and execution of this protocol, please refer to Krieg et al.1.