Amyloid-beta plagues and neurofibrillary tangles are key pathological features observed in the brains of Alzheimers patients. Familial, early onset forms of AD (FAD) are caused by autosomal dominant inherited genetic mutations and offer an opportunity to study the effects of key mutations on the diseases progression and pathology. To date, approximately 200 FAD mutations in APP and/or PSEN1 have been reported. Transgenic mouse models and human neurons harboring FAD mutations in PSEN-1 and/or APP are widely used as model systems and have provide important insights into the disease. Transgenic mouse models are able to recapitulate the loss in cognitive functions along with increased deposition of b-amyloid plagues, but are unable to demonstrate neurofibrillary tangles, one of the key pathological hallmarks of AD. Similarly human iPSC-derived neurons from FAD patients demonstrated increased levels of the pathogenic AB species and phosphorylated tau, but lack the characteristic amyloid-beta plagues and neurofibrillary tangles.Recently a 3D model using genetically engineered human neural stem cells that overexpress FAD mutations was reported to recapitulate the two pathological hallmarks of AD: b-amyloid plagues and neurofibrillary tangles. Lentiviruses expressing FAD mutations in human APP with both the K670N/M671L (Swedish) and V717I (London) mutations (APPSL) and/or PSEN1 with the E9 mutation (PSEN1(E9)) and APPSL/PSEN1(E9) along with fluorescent proteins as reporters for viral infection (see below), were used to transfect ReNcell VM human neural stem cells. FACS was utilized to enrich for cells with the highest transgene expressions followed by encapsulation of the sorted cells in a 3D Matrigel culture system. After approximately 6 weeks of differentiation, aggregation of AB was observed. Robust accumulation of phosphorylated tau along neurofibrillary tangles was readily detectable after 10-14 weeks.The Alzheimers In A Dish PSEN1-RFP Lentivirus is a necessary reagent to set up the Alzheimers 3D culture system. The lentivirus contains the PSEN1 FAD mutations and can be used to infect human neural cells. The RFP tag enables assessment of viral infectivity and allows FACS sorting of the highest expressing cells.
Synonyms: PSEN1-RFP, ReNcell 3D Alzheimers Model, ADID 3D Neural Cell Model
Application: Research CategoryStem Cell ResearchNeuroscience
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