A 3D human neural cell culture system for modeling Alzheimer's disease

Developers

Young Hye Kim, Se Hoon Choi, Carla D’Avanzo, Rudolph E. Tanzi, Doo Yeon Kim, etc.

Description of the technology

Stem cell methods 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, because existing animal models have been unable fully to recapitulate all aspects of pathology.

The technology is based on an improved version of novel 3D cell culture system that exhibits key events in Alzheimer’s disease pathogenesis, including extracellular aggregation of amyloid-β and accumulation of hyperphosphorylated tau protein. The basic version of this 3-D cell culture includes genetically engineered human neural stem cells that overexpress genes, related autosomal-dominant forms of Alzheimer’s disease, combined with a 3D culture conditions. In process of modernization of basic version of 3D human neural cell culture system for modeling Alzheimer’s disease authors of the technology have developed universal instructions for the generation and analysis of these 3D human neural cell cultures. These instructions describe the production of genetically modified human neural progenitor cells (hNPCs) with mutations of familial Alzheimer’s disease, the differentiation of the hNPCs in a 3D matrix and the analysis of Alzheimer’s disease pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of amyloid-β is observed after 6 weeks of differentiation, followed by robust tau protein pathology after 10–14 weeks. The feature of this 3D culture is that it uses a special cultural medium — Matrigel. Therefore the technology has an advantage, because in 2D models, secreted amyloid-β may diffuse into the cell culture medium, disrupting aggregation, whereas the 3D Matrigel may prevent this diffusion of amyloid-β, allowing for high local concentrations that are sufficient to initiate aggregation. Thus, 3В cell culture allows to develop robust and stable model of Alzheimer’s disease.

Practical application

This protocol can be used to examine many central characteristics of Alzheimer’s disease pathogenesis in vitro, including the molecular mechanisms underlying the production of high concentrations of amyloid-β, the accumulation of extracellular amyloid-β, the deposition of amyloid-β aggregates, the hyperphosphorylation of tau protein aggregation. These paths may lead to new diagnostic and prognostic biomarkers of Alzheimer’s disease The model can also be used to test other genetic or environmental factors associated with Alzheimer’s disease in conjunction with the mutations, causing autosomal-dominant forms of Alzheimer’s disease. The flexible scalability of the system makes it applicable for use in larger-scale testing and drug screening. This protocol may also be useful to other neurodegenerative diseases with genetic variations, and it may be especially suitable for diseases characterized by abnormal aggregation of misfolded proteins.

Laboratories

• Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown (USA)
• Biomedical Omics Group, Korea Basic Science Institute, Cheongju-si, Chungbuk (Republic of Korea)
• Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn and Hertie Foundation, Bonn (Germany)

Links

Publications

• Kim, Y.H. et al. «A 3D human neural cell culture system for modeling Alzheimer’s disease." 10 Nature Protocols, (2015): 985–1006.
• Choi, S.H. & Tanzi, R.E. «iPSCs to the rescue in Alzheimer’s research." 10 Cell Stem Cell, (2012): 235–236.
• Choi, S.H. et al. «A three-dimensional human neural cell culture model of Alzheimer’s disease." 515 Nature, (2014): 274–278.