The experimental procedure begins with the generation of human neural progenitor cell lines virally transfected with APP and/or PSEN1 FAD mutations and enriched based on GFP and/or mCherry signals by fluorescence-activated cell sorting (FACS). The right-hand column shows in detail the timing for each step of the 3D culture method. Day 0 indicates the day on which cells are mixed with Matrigel. MAP2, microtubule associated protein 2; 4R tau, 4-repeat tau isoform; vGluT1, Vesicular glutamate transporter 1; NR2B, N-methyl D- aspartate receptor subtype 2B; TH, Tyrosine Hydroxylase.