Protocol to generate murine organotypic brain cultures for drug screening and evaluation of anti-metastatic efficacy

Summary: Organotypic brain cultures are short-term assays that phenotypically and functionally recapitulate brain metastatic cells in vivo. Here, we present a protocol to generate murine organotypic brain cultures for drug screening. We describe steps for sectioning of murine brains and plating of organotypic cultures. We then detail evaluation of the anti-metastatic effect of chemical compounds through bioluminescence imaging before and after drug treatment. Combined with downstream applications, this protocol allows comprehensive characterizations of both cancer cells and the tumor-associated microenvironment.For complete details on the use and execution of this protocol, please refer to Zhu et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.