A living biobank of ovarian cancer ex vivo models reveals profound mitotic heterogeneity

High-grade serous ovarian carcinoma is characterised by TP53 mutation and extensive chromosome instability (CIN). Because our understanding of CIN mechanisms is based largely on analysing established cell lines, we developed a workflow for generating ex vivo cultures from patient biopsies to provide models that support interrogation of CIN mechanisms in cells not extensively cultured in vitro. Here, we describe a “living biobank” of ovarian cancer models with extensive replicative capacity, derived from both ascites and solid biopsies. Fifteen models are characterised by p53 profiling, exome sequencing and transcriptomics, and karyotyped using single-cell whole-genome sequencing. Time-lapse microscopy reveals catastrophic and highly heterogeneous mitoses, suggesting that analysis of established cell lines probably underestimates mitotic dysfunction in advanced human cancers. Drug profiling reveals cisplatin sensitivities consistent with patient responses, demonstrating that this workflow has potential to generate personalized avatars with advantages over current pre-clinical models and the potential to guide clinical decision making.


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All studies must disclose on these points even when the disclosure is negative.  MTAB-7225, E-MTAB-7223, E-MTAB-724, E-MTAB-8559 and PRJEB28664 respectively. The data underlying Figs. 2b, 3a, d, 4b, c, d, e, g, 5b, e, 6b, 7a, b, d, 8b, c, 9b, c, 10a, d, S2a, S4a, b are provided as a Source Data file. All other data supporting the findings of this study are available within the article, the Supplementary information files, or the corresponding author upon request. A reporting summary for this article is available as a Supplementary Information file.
We describe 15 ex vivo cultures generated from 12 patients as a proof of principle cohort. Beyond that, where appropriate, sample sizes are denoted in the figure legends.
During the study we realized that OCM69 was a stromal culture; rather than exclude it, we retained it to provide an additional internal control for the NGS experiments. If it became apparent that a stromal culture underwent senescence during an experiment, it was excluded and the experiment repeated with an earlier passage.
Descriptive analyses were confirmed via orthogonal approaches: e.g. TP53 mutations were identified by Sanger sequencing of RT-PCR products, exome sequencing and targeted amplicon sequencing. Doubling times and IC50 values were determined by analyzing three technical replicates and at least three biological replicates.
There was no need to randomize samples in this study.
For the cell biology studies the samples were not blinded. For the Exome, RNAseq, scRNAseq, scWGS, M-FISH and IHC, samples were processed and analyzed by colleagues without specifically knowing what any given sample represented.