A HUMAN 3D NEURAL CULTURE MODEL UTILISING INDUCED PLURIPOTENT STEM CELLS FROM AN ALZHEIMER’S PATIENT TO STUDY MECHANISMS OF ALZHEIMER DISEASE PATHOLOGY

Abstract

Alzheimer's Disease (AD) is the leading cause of dementia and with a population of increasing longevity is set to become a major burden on health services. Currently human tissue from AD cases is only available for research from post mortem brain but we can now utilise iPSC technology to obtain neural stem cells from AD patients with the aim to produce mature neurons with the same genetic signature of AD patients. Using induced Pluripotent Stem Cells (iPSCs) from early onset AD patients containing Presenilin 1 mutations we have created a 3D human neural culture system for the study of some early onset Alzheimer's disease mechanisms. Using immunofluorescence, western blotting and electrophysiology we have characterised neuronal morphology and maturity, synaptic function and disease-protein profiles. Neural progenitors derived from AD iPSCs (Axol Biosciences) were plated in matrigel and self-organised into 3D structures by week 3. These cultures contained a heterogeneous cell population expressing differentiation markers for neurons and astrocytes and importantly AD-associated proteins, amyloid precursor protein/Abeta and tau. These 3D cultures were maintained in vitrofor 18+ weeks. They expressed the mature 4R tau isoform and Abeta oligomers by week 6 and exhibited repetitive firing by week 12. Together these results suggest the presence of mature neurons within these cultures and the expression of pathology-associated proteins. We have created 3D mature neural cultures derived from AD patients to study mechanisms of the disease pathology in a human cell model. This is a viable new in vitro 3D neural model for the study of neuropathology, which can serve as a powerful new translational tool in this area of research.