Journal: Journal of Virology
Article Title: Macrophages derived from human induced pluripotent stem cells (iPSCs) serve as a high-fidelity cellular model for investigating HIV-1, dengue, and influenza viruses
doi: 10.1128/jvi.01563-23
Figure Lengend Snippet: Blood- and iPSC-derived macrophages are phenotypically similar. (A) iPSCs were differentiated into monocytes using the STEMdiff monocyte kit (STEMCELL Technologies) (top row bright-field images). As controls, blood was drawn from healthy donors and monocytes were isolated through a series of density-gradient purifications (bottom row bright-field images; see Materials and Methods). Both blood- and iPSC-derived monocytes were differentiated into naïve M0 macrophages via 10-ng/mL macrophage colony stimulating factor (M-CSF) treatment for 4 days. Naïve macrophages were then polarized into M1 macrophages using 50-ng/mL interferon gamma (IFN-γ) and 10-ng/mL lipopolysaccharide (LPS), or into M2 macrophages using 10-ng/mL interleukin (IL)-4, with either treatment lasting 48 hours. Scale bars = 40 μm. (B and C) Surface protein expression levels were quantified via fluorescent antibody staining followed by flow cytometry (one representative experiment shown from three biological replicates). Monocytes and the subsequent macrophage subtypes derived from blood and two patient-derived iPSC lines were used. (B) Expression levels of monocyte markers CD14 and CD11b were quantified on freshly isolated monocytes. (C) Following macrophage polarization, expression levels for CD80 (M1 macrophage marker) and CD206 (M2 macrophage marker) were quantified within the CD14+ macrophage subset.
Article Snippet: Monocytes were differentiated from iPSCs using the STEMdiff Monocyte Kit (STEMCELL Technologies, #05320), following the manufacturer recommendations.
Techniques: Derivative Assay, Isolation, Expressing, Staining, Flow Cytometry, Marker
