Assessment of Tau Aggregation in Human, iPSC-derived Neurons

A client requested a custom, neuronal model to evaluate their therapeutic agent’s effects on tau expression and phosphorylation using iPSC-derived glutamatergic neurons from two different donor backgrounds.

Goals:

• Establish an assay to measure tau expression and phosphorylation in healthy and MAPT P301S human, iPSC-derived glutamatergic neurons.
• Evaluate the effects of client’s therapeutic agent on various measures of tau in each culture model.

Cell Model

iPSC-glutamatergic neurons

• WT/WT
• MAPT P301S

Palette 1

Hoechst (nuclei)
Tuj1
Total Tau
Phospho-tau Ser202/Thr205 (AT8)

Palette 2

Hoechst
Tuj1
Total Tau
Phospho-tau Thr217 (6G7C8)

Treatments and Timelines

• Culture cells in 384-well, imaging microplates, following custom culture protocol developed at PhenoVista until DIV12.
• On DIV12, treat with test articles
• Continue with half-media changes every other day for two weeks, readministering treatments with each change.
• On DIV26, treat appropriate control wells with okadaic acid (inhibitor of serine/threonine protein phosphatases, causing tau hyperphosphorylation) 3hrs prior to fixation.
• Fix and stain cultures for scanning and analysis.

REPRESENTATIVE IMAGES & QUANTITATIVE DATA