Alzheimer's disease (AD) is an age-related neurodegenerative disease that leads to insidious loss of neurons, and subsequently to dementia. Amyloid-b (Ab) and tau are the two pathological hallmarks, and both are known to be dysregulated in AD leading to widespread proteinaceous aggregation in the brain. We now know from tau-deficient animal models that tau does not merely facilitate Ab-induced toxicity but is also essential in that particular signalling cascade. However, a tau-deficient human in vitro model has not been available to corroborate the Ab-tau interaction. Here we report generation of the first MAPT knockout (tau-KO) human induced pluripotent stem cell (iPSC) lines in order to study Ab-tau interaction in human biological context. We also characterised a versatile and scalable cortical neuron differentiation protocol that successfully produced a heterogeneous population of functional neurons manifesting cortical identity. Furthermore, we extracted brain homogenate from AD patient to serve as a source of Ab. Application of brain homogenate, which contains soluble Ab in pM range, in iPSC-derived neuronal culture resulted in neurite degeneration and cytotoxicity within 72 hours. We then asked if the human tau-KO genetic background could exhibit neuroprotective effects upon Ab exposure to the neuronal culture. We have initiated an investigation into the interaction between Ab and tau in human iPSC tau-KO cortical neurons utilising patient brain homogenate as the source of Ab to mimic human AD pathology in vitro.