Journal: Cell Reports Methods

Article Title: Cortical organoid-derived models of the melanoma brain metastatic niche enable prioritization of cancer-targeting drugs

doi: 10.1016/j.crmeth.2025.101236

Figure Lengend Snippet: Anti-cancer drug screen and MBM validation strategy (A) Schematic overview of the drug repurposing screening strategy starting with the primary screen using 2D A375 cells and MelDCCs from patient’s lymph node specimen. (B) Flow cytometry analysis of MelDCC and A375 cells stained for Melan-A and NGFR receptor. (C) Heatmap of ATP-based cellular viability of A375, MelDCC3, MelDCC8, and MelDCC11 cells and NHEM control cell line after 5-day exposure to 315 anti-cancer drugs screened at 1 μM. Viability values are ranked from left to right based on their impact on the mean viability of all cell lines. Each data point is represented by one individual colored line. (D) Workflow outlining the criteria and steps for rational hit selection starting from 2D screening data. (E) Close-up of heatmap showing the sensitivity of the cell panel including neural progenitor cells (NPCs) toward selected compound hits. (F) Concentration-response titration of selinexor on ATP-based cellular viability of 2D and tumoroid (3D) cell cultures ( n = 2–3 experiments per condition). Data are presented as mean ± SEM. IC 50 (2D MelDCC3): 0.83 μM; IC 50 (3D MelDCC3): 0.54 μM; IC 50 (2D MelDCC8): 0.6 μM; IC 50 (3D MelDCC8): 0.4 μM. (G) Fluorescence-based cellular viability of A375-MBMs treated with hit compounds at 10 μM for 72 h ( n = 3–5 experiments). Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA, followed by Tukey’s multiple comparisons test. ∗ p = 0.0179. See also .

Article Snippet: Intracellular marker Melan-A (1:40, Santa Cruz Biotechnology, Sc-20032) or isotype control (1:20, BioLegend, 400155) was stained for 30 min at room temperature.

Techniques: Biomarker Discovery, Flow Cytometry, Staining, Control, Selection, Concentration Assay, Titration, Fluorescence