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. 2017 Feb 6;36(25):3576–3587. doi: 10.1038/onc.2016.509

Figure 1.

Figure 1

Stromal fibroblasts enhance colony formation of metastatic ovarian cancer cells in three-dimensional (3D) organoid model with de-novo expression of 16 cytokines. (a) Schematic representation of 3D organoid co-culture model (left, upper panel). Normal stromal fibroblasts (WI38) were mixed with extracellular matrix (ECM; BD Matrigel™) and placed at the bottom of chamber slides. Single-cell suspension of metastatic ovarian cancer cells (SKOV3) was then overlaid on top of ECM with WI38. Monoculture of SKOV3 with ECM alone was served as controls. The data represents means±s.e.m. from three independent experiments. ***P<0.001, *P<0.05 (Mann-Whitney U test). (b) Conditioned medium (CM) from ECM alone (Matrigel), 3D monoculture of WI38, 3D monoculture of SKOV3, and 3D organoid co-culture model (SKOV3+WI38) were collected on day 4 (left, upper panel) and their relative protein expressions of a total of 174 cytokines were compared using cytokine antibody array (RayBiotech AAH-CYT-G2000-4). The signal was then normalized by the internal positive control and subject to clustering analysis using DChip software (left, lower panel). A panel of cancer-specific cytokines was identified in the CM of 3D organoid monoculture of SKOV3 ovarian cancer cells (right, upper panel). TNF-α was indicated by a red arrow. De-novo expression of 16 cytokines was identified in the CM of 3D organoid co-culture (right, lower panel). TGF-α was indicated by a red arrow. (Blue: low expression level; White: no change; Red: high expression level).