Published June 16, 2023 | Version v1
Publication Open

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

Authors/Creators

  • 1. ROR icon Spanish National Cancer Research Centre

Description

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.

Technical info

We want to thank I. Blanco for critical review of the manuscript. We also thank J. Massague (MSKCC) for the H2030-BrM cell line. This work was supported by MINECO (SAF2017-89643-R, SAF2014-57243-R, SAF2015-62547-ERC) (M.V.), Fundacion FERO (IX FERO Grant for Research in Oncology) (M.V.), Fundacio La Marato de TV3 (141) (M.V.), Melanoma Research Alliance (Bristol-Myers Squibb-Melanoma Research Alliance Young Investigator Award 2017 [https://doi.org/10.48050/pc.gr.75716]) (M.V.), Beug Foundation (Prize for Metastasis Research 2017) (M.V.), Fundacion Ramon Areces (CIVP19S8163) (M.V.), Worldwide Cancer Research (19-0177) (M.V.), H2020-FETOPEN (828972) (M.V.), Cancer Research Institute (Clinic and Laboratory Integration Program CRI Award 2018 (54545)) (M.V.), AECC (Coordinated Translational Groups 2017 (GCTRA16015SEOA) (M.V.), LAB AECC 2019 (LABAE19002-VALI) (M.V.), ERC CoG (864759) (M.V.), MICIN (PID2021-124582OB-I00) (M.V.), ASPIRE Award, The Mark Foundation (ID: 21-05-ASP), La Caixa-Severo Ochoa International PhD Program Fellowship (LCF/BQ/SO16/52270014) (L.Z.), and Boehringer Ingelheim Fonds MD fellowship (L.M.). CNIO is supported by the ISCIII, the Ministerio de Ciencia e Innovacion and is a Severo Ochoa Center of Excellence (SEV-2015-0510). M.V. was named Ramon y Cajal Investigator (RYC-2013-13365) and is a member of EMBO YIP (4053).

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Additional details

Funding

European Commission
NanoBRIGHT - BRInGing nano-pHoTonics into the brain 828972
European Commission
ALTER-brain - Metastasis-associated altered molecular patterns in the brain 864759

Dates

Accepted
2023-06-16