Amyloid-β42/40 ratio drives tau pathology in 3D human neural cell culture models of Alzheimer’s disease

Clonal FAD hNPCs generated by single cell sorting

Heterogeneous FAD hNPCs were generated as previously described by expressing lentiviral DNA constructs harboring APPSL and/or PS1ΔE9 (Supplementary Table 1)^21,22. To obtain clonal FAD hNPCs, heterogeneous FAD hNPCs were sorted into single cells using a FACS-assisted single cell sorting protocol based on GFP and/or mCherry signals (Fig. 1a, b and Supplementary Fig. 1). Viable cells occupying individual wells in 96-well plates were expanded to form colonies within 2–3 weeks (Fig. 1c). For slow growing colonies, we added naïve ReNcell VM cells to accelerate cell proliferation. Then, hNPCs with GFP and/or mCherry expressions were enriched again by an additional FACS-assisted bulk cell enrichment protocol (Fig. 1a). Next, we analyzed secreted Aβ levels of clonal control and FAD hNPCs by Western blot analysis. As expected,…