The development of new compounds capable of inhibiting or preventing tau uptake and seeding is moving in the focus of Alzheimer’s disease treatment. Consequently, reliable in vitro models that mirror this tau pathology in human diseases are needed. We established and optimized tau uptake assays using a stably transfected tau overexpressing SH-SY5Y cell line and mouse primary cortical neurons in combination with human AD seeds.
Figure: Tau uptake (A,C) and seeding (B,D) in stably transfected SH-SY5Y cells overexpressing 2N4R tau with P301L mutation (A, B) and mouse primary cortical neurons (C,D). Cells were incubated for 48 h with tau seeds extracted from AD brains (AD seeds) or the same seeds, co-incubated with an anti-tau antibody (HT7) as reference item to reduce seeding and uptake. Tau seeding was assessed by application of 2.5 µg seeds (total protein) in the presence of lipofectamine to the cells, tau uptake was monitored by applying 20 µg seeds (total protein) in the absence of lipofectamine. Tau aggregation was assessed 48 h after application of the seeds using HTRF-based Tau Aggregation Kit from Cisbio. Vehicle treated cells served as control. One-Way ANOVA with Dunnett’s multiple comparisons test vs. vehicle control (VC). Mean + SEM; n = 6. ***p<0.001.