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. 2020 Sep 15;11:4629. doi: 10.1038/s41467-020-18388-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Workflow used to set up a RMS pre-clinical drug profiling platform. Briefly, PDXs were established from small RMS biopsies and first enzymatically dissociated to derive PPCs, followed by a culture condition screen (see text). DNA-sequencing and aCGH analysis were performed on both PDXs and paired PPCs to generate a sample collection with annotated genomic information. Finally, a high-throughput drug screen was conducted on each PPC cultivated under optimal culture conditions and top drug candidates were further validated in vivo on the original PDXs. b Culture optimization of indicated PPCs. Heat map depicting cell viability scores (n = 2–5 biological replicates), differentiation status (n = 2–3 biological replicates) and quantification of contamination with mouse stroma cells (n = 2–3 biological replicates) after short-term culture (1–3 weeks) of PPCs is shown. nd, not determined due to either low number of surviving cells or high fraction of cell loss during washing steps. c Growth curves of indicated PPCs cultured in DMEM on uncoated plates (black line) or in NB + GF on matrigel (red line). Cell number was normalized to day of seeding and is expressed in a log2 scale. (Mean ± range; n = 2 biological replicates). d Graph summarizing the proliferation rate of 11 PPCs under optimal and standard DMEM conditions. Data express the percentage of cells after 5 passages relative to day 0 in a log2 scale (n = 2–3 biological replicates). The dotted line indicates the cell number at day 0. e, left panel, Viability of the human cell fraction in PPCs from RMS-AMS007 PDXs cultured under indicated conditions (Mean ± range; n = 2 biological replicates). e, right panel, Viability of PPCs from RMS-AMS007 cultured in indicated media supplemented with ROCK- (Y-27632) and/or TGFβ- (A83-01) inhibitors (Mean ± sd; n = 3 biological replicates; two-way ANOVA with Tukey’s multiple comparisons test). f, left panel, Tumor growth kinetics of RMS-ZH004 PDX (blue, n = 3 biological replicates) and passage 4 CDX (green, n = 5 biological replicates). (Mean ± sem). f, right panel, Histogram indicating the day (d) of sacrifice of individual CDX- and PDX-bearing mice (gray dots) when tumors reached ~1000 mm³. (Mean ± sd; two-tailed paired t-test; NS, not significant;). Source data are provided as source data file.