HUMAN INDUCED PLURIPOTENT STEM CELL-BASED APPROACH FOR THE STUDY OF ALZHEIMER’S DISEASE PATHOLOGHY AND NEUROINFLAMMATION
Alzheimer’s disease (AD) is a progressive, neurodegenerative disorder and the most prevalent cause of dementia in elderly people. Many molecular events contributing to the formation and progression of Alzheimer’s disease have been uncovered and carefully investigated. However, the precise contribution and succession of these events remain elusive. Three major contributors to AD pathology are an accumulation of pathological forms of amyloid-β protein (amyloid β1-40 and β1-42), the formation of intraneuronal neurofibrillary tangles composed of the abnormal protein tau and pronounced neuroinflammation. Progressive nature of these pathologies together finally leads to neurodegeneration, cognitive decline and eventually death of the patient. An important element of the brain’s cellular composition is microglia – the tissue-specific ‘macrophage’ of the brain parenchyma. This cell type highly contributes to the pro-inflammatory environment of the AD brain by mechanisms such as cytokine detection and ATP-signalisation (e.g. per P2X7 receptor activation). Here, we present the generation of iPSC-derived microglial cells through a 3D differentiation protocol, using bone morphogenetic protein 4 (BMP4), vascular endothelial growth factor (VEGF), and stem cell factor (SCF). Further differentiation was forced by adding macrophage colony-stimulating factor (M-CSF) and interleukin-3 (IL-3). The acquired cells express microglial markers such as Iba1, CX3CR1, CD11c, and TMEM119, which confirms their suitability for further use in a mixed culture with iPSC-derived neural cells. Using this mixed cellular model, we will be able to investigate in vitro the role of microglia in the development and progression of AD, with special focus on the role of the P2X7 receptor and its activation.