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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: Ovarian cancer stem cells and macrophages reciprocally interact through the WNT pathway to promote pro-tumoral and malignant phenotypes in 3D engineered microenvironments

Fig. 1

Hetero-spheroids derived from monocyte-derived macrophages and ovarian cancer stem cells. a Monocytes from the U937 cell line or peripheral blood monocytes (PBMC) were plated into hanging drop arrays, and differentiated to M0 macrophages with phorbol ester treatment, or activated with IL-4 and MCSF treatment. Differentiated and activated macrophages formed compact spheroid-like aggregates. b Differentiated M0 and M2 macrophages all expressed pan macrophage marker, CD68 indicating 75–80% differentiation efficiency from monocytes to macrophages. CD68 expression was evaluated using flow cytometry analysis, with representative plots. c Polarization was assessed using qPCR analysis for two M2 genes, CD163 and CD206. Both U937 and PBMC macrophages expressed significantly higher levels of CD163 and CD206 genes, compared to untreated undifferentiated monocytes. d M2 differentiated macrophages secreted higher amounts of the immuno-suppressive cytokine IL-10, and the pro-tumoral cytokine IL-6. e CSCs were derived from the OVCAR3 cell line based on ALDH+ CD133+ expression. Hetero-spheroids were generated using differentiated U937 or PBMC M0 macrophages and CSCs or activated U937 or PBMC M2 macrophages and CSCs. Representative phase contrast images of hetero-spheroids seen at Day 5 following formation indicate compact spheroids, similar in size to CSC mono-spheroids generated from the same number of CSCs/spheroid. f Hetero-spheroids retain ~ 20% CD68 expression, indicating that at day 5, CD68+ macrophages constitute 20% of the population of cells. Scale bar = 200 μm

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