Fig. 2: Individual clonal populations fail to form solid peritoneal metastases.

a Representative images of OCI-C5x generated malignant ascites, and b solid tumors on the diaphragm, which connect to the liver (white arrow). c H&E-stained sections from the solid tumor masses, representative images from one of three independent experiments. Note that the tumor cells maintain histopathological characteristics of CCC in patients with “clear” cells (C-left image, arrow) and hobnail features (C-right image, arrow). Scale bar, 100 μm. d Representative H&E images of tumor sections of solid metastases on diaphragm generated by CL31, CL49, and the multiclonal mixture, representative images from one of two independent experiments. T tumor cells, D diaphragm, L liver. Scale bar, 5 mm. e Tumor burden score of solid metastases on the diaphragm (data from two independent experiments, n = 8–10 mice per group). Scored blindly. p Values were computed using the chi-square test with Monte Carlo simulation. f Schematic depiction of experimental design and workflow for barcode experiments. g Barcode representation in the indicated samples collected from mice injected with the multiclonal mixture composed of barcoded clones, and h in samples collected from monolayer and suspension cultures in vitro. A representative of two independent experiments.