Genetically Engineered Macrophages Derived from iPSCs for Self-Regulating Delivery of Anti-Inflammatory Biologic Drugs

Figure 3

Polarization and signaling in response to cytokine treatment with either no treatment (NT), IFNγ/LPS, or IL-4/IL-13 stimulus after 24 hours. (a) Phase contrast imaging of iMAC in comparison to BMDMs. (b) qPCR normalized to GAPDH demonstrated similar upregulated gene expression in response to inflammatory or immunomodulatory stimuli, with lower overall fold-change response for iMACs than in BMDMs. Heat map color represents downregulation (blue), upregulation (red), or no change (grey) per gene (n = 6). (c) Western blot analysis of iMACs/BMDMs cytokine activation. (d) Griess assay for nitric oxide production of iMAC/BMDMs in response to inflammatory stimuli. (e) Phagocytosis in iMACs/BMDMs quantified by flow cytometry. (f) Migration images from Boyden chamber for iMACs/BMDMs, naïve, or prestimulated with IFNγ/LPS, in response to chemoattractants (n = 4). (d, e) Bars represent mean ± SEM with # (iMAC) or (BMDM) denoting to no treatment by one-way ANOVA.