Fig. 2: Dissociation of tumor tissue workflow, DST analysis and validation of patient-derived tumor cultures.

a, Tissue processing and derivation of short-term PDCs, including representative images of received tissues (left) and derived PDCs (right) from EV004-RMS, EV007-GBM, EV010-EWS and EV014-MRT. b, Ex vivo DST using a library of more than 125 FDA-approved drugs, post-endpoint quality control process based on Z-prime scores, IC₅₀ and DSS analysis, and representative results from single agent testing for EV010-EWS followed by physician-selected drug combinations (if additional PDC material remained). Lum, luminescence. * indicates physician feedback guided selection of tested drug combinations. The slim red borders around single agents on the left indicate those included in combination testing, The thick red border on the right indicates the final drug combination used for the patient. c,d, Molecular characterization and validations of PDCs assessed by immunofluorescence detection of pathology-defined markers in EV010-EWS (c) and EV019-MB (d). Immunofluorescence images of one independent experiment (due to limited PDC material). e,f, Analysis of RT–qPCR to confirm loss of TP53 transcripts in EV003-OS (e) and DIS3L2 transcripts in EV015-WT (f). g,h, Immune cell type deconvolution and tumor purity analysis from tumor tissue at enrollment (T) and PDC in EV004-RMS (g) and EV009-OS (h) using bulk RNA-seq deconvolution tools EPIC, ESTIMATE and quanTIseq (right panel). Representative pie charts present EPIC deconvolution results. TC, tumor cell. Portions of panels a and b were created with BioRender.com.