To ascertain whether the AD phenotypes of Aβ aggregation and Tau hyperphosphorylation are generalizable to fAD organoids from different sources, rather than being a phenomenon of the APPDp1-1 line, we examined additional organoids created from patient-derived and healthy control iPSCs (S1 Table). At 90d of culture, we processed and analyzed organoids created from an additional APP duplication line (APPDp2-3; [44]) two PSEN1 fAD mutant lines (PSEN1M146I, PSEN1A264E; Fig 2A, S1 Table). Organoids derived from fAD patients with APP duplication (APPDp2-3) or the PSEN1A264E mutation exhibited increased numbers of Aβ aggregates verses the control lines (Fig 2B). While the fAD organoids from PSEN1M146I patient cells exhibited a trend towards higher amyloid levels, this was not significant. We also examined organoids from these additional fAD lines for the presence of pTau immunoreactivity. Similar to the pattern observed with amyloid aggregation, the APPDp2-3 and the PSEN1A264E fAD organoids exhibited increased pTau (Ser396) immunoreactivity at 90 days of culture, while the PSEN1M146I organoids did not differ from control (Fig 2C).
