Kim, Young HyeChoi, Se HoonD'Avanzo, CarlaHebisch, MatthiasSliwinski, ChristopherBylykbashi, EnjanaWashicosky, Kevin J.Klee, Justin B.Brüstle, OliverTanzi, RudolphKim, Doo Yeon2022-07-152015-06-11Kim, Young Hye, Se Hoon Choi, Carla D'Avanzo, Matthias Hebisch, Christopher Sliwinski, Enjana Bylykbashi, Kevin J. Washicosky et al. "A 3D Human Neural Cell Culture System for Modeling Alzheimer's Disease." Nature Protocols 10, no. 7 (2015): 985-1006. DOI: 10.1038/nprot.2015.0651754-21891750-2799https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37372637Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer’s disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks.en-USResearch Subject Categories::MEDICINE::Morphology, cell biology, pathology::Cell biology::NeuroscienceJournal Article2022-07-1510.1038/nprot.2015.065