3D co-culture polarized MDMs resemble M2-like macrophages.

<p>Tumor cells and fibroblasts were co-cultured for 5 days. Monocytes were added to co-culture on day 5 to differentiate for 6 days. Spheroids were collected and dissociated by using Accutase to obtain a single cell suspension. A) Single cell suspensions were analysed by flow cytometry using the illustrated gating strategy for MDM phenotyping. B) Cell surface marker expression of 3D myeloid cells was compared to <i>in vitro</i> generated M2c and activated M1 macrophages. Typical M2 and M1 macrophage marker were analyzed by flow cytometry. 3D co-culture MDMs expressed high levels of CD163 and CD14 and low levels of CD86 and HLA-DR comparable to <i>in vitro</i> differentiated M2 macrophages (dotted box). Shown is one representative out of n = 5 experiments, red numbers show geometrical mean values for relevant markers.