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

Figure 4

(a) ELISA for sTNFR1 in response to varying levels of TNF in Ccl2-sTNFR1 iMACs 72 hours following treatment (n = 3–6). (b) qPCR normalized to GAPDH in response to treatment with TNF-α after 24 and 72 hours in sTNFR1 edited iMACs (n = 3–5). (c) Ccl2-Luc iMAC luciferin production in response to TNF-α compared to no treatment (n = 3). (d) ELISA for sTNFR1 in response to TNF-α in Luc/sTNFR1 iMACs (n = 3-4). (e) qPCR normalized to GAPDH in response to treatment with TNF-α after 24 and 72 hours in sTNFR1 edited iMACs (n = 3–5). (f) ELISA over time for sTNFR1 iMACs in response to repeated stimulations as shown by red arrows (media change with either no treatment or 20 ng/mL TNF-α) in sTNFR1 iMACs (n = 3). (g) qPCR normalized to GAPDH and an untreated control in response to treatment with IFNγ/LPS or IL-4/IL-13 after 24 hours or to TNF-α after 24 and 48 hours (n = 3). (h) Griess assay for nitric oxide production in response to inflammatory stimuli (n = 4–8). (i) Phagocytosis in nontreated Ccl2-Luc and Ccl2-sTNFR1 iMACs quantified by flow cytometry, and technical replicates represent 50,000 events. Bars represent mean ± SEM, , , , and by one-way ANOVA. NT: no treatment.