Differentially polarized human monocyte-derived macrophages (M0, M1, M2/IL10, M2/IL4, TAM-like)

In order to assess whether differences in the transcriptomic profile of human tumor-associated macrophages vs. alveolar macrophages from adjacent non-tumor-tissue (GSE162669) were caused by factors secreted by tumor cells, primary human monocyte-derived macrophages (MDMs) were polarized towards a TAM-like phenotype by cultivating them in tumor cell-conditioned medium. A genome-wide comparison by bulk RNA-Seq confirmed a high similarity of ex vivo TAMs and in vitro polarized TAM-like macrophages. Other polarization schemes (M1: LPS/IFN-gamma, M2: IL10 or IL4) did not lead to similar transcriptomic changes. For details, see Hoppstädter et al., 2021 (doi:10.1016/j.ebiom.2021.103578). Overall design: After differentiation, MDMs were polarized for another 24 hours. To this end, MDM medium was supplemented with 20 ng/ml IFN? and 100 ng/ml LPS for M1 polarization; either 20 ng/ml IL4 or IL10 for M2 polarization; or left without further supplementation for M0 macrophages. TAM-like macrophages were generated by cultivation in A549 tumor cell-conditioned medium (TCM) supplemented with 20 ng/ml M-CSF. MDM Samples from one female (donor 1) and two male donors (donor 2 and 3) were used for RNA sequencing.