Characterization of iPSC-derived dopaminergic neurons, microglia, and co-cultures. Results are shown for three iPSC lines derived from healthy individuals (SFC086-03-03, SFC089-03-07, and SCF156-03-01). (A) Differentiation schemes are shown for microglial (MG) and dopaminergic neuronal (DAN) monocultures, and a combined co-culture (co-DAN-MG). Protocols are based on published methods [15,16,17,18,19]. (B) Flow cytometry analysis of the surface markers cluster of differentiation (CD) 14, 45, and 88 in myeloid precursor cells (purple) and iPSC (grey). Images are representative of n = 10 independent differentiation experiments. (C) Immunofluorescence staining for microglial ionized calcium-binding adaptor molecule 1 (IBA1) (green), CD88 (green), neuron-specific β-III tubulin (TUJ1) (red), and the dopaminergic marker tyrosine hydroxylase (TH) (cyan). Images are representative of n = 10 independent differentiation experiments. Scale bar, 25 μm. (D) Western blot analysis of IBA1 and TH marker proteins and loading control GAPDH in the three different cultures derived from the three iPSC lines. Whole blots are shown in Figure S4.