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

被引：193

作者：

Kim, Young Hye ^{[1
,2
]}

Choi, Se Hoon ^{[1
]}

D'Avanzo, Carla ^{[1
]}

Hebisch, Matthias ^{[1
,3
,4
]}

Sliwinski, Christopher ^{[1
]}

Bylykbashi, Enjana ^{[1
]}

Washicosky, Kevin J. ^{[1
]}

Klee, Justin B. ^{[1
]}

Bruestle, Oliver ^{[3
,4
]}

Tanzi, Rudolph E. ^{[1
]}

Kim, Doo Yeon ^{[1
]}

机构：

[1] Harvard Univ, Sch Med, Massachusetts Gen Hosp, MassGeneral Inst Neurodegenerat Dis,Genet & Aging, Charlestown, MA 02129 USA

[2] Korea Basic Sci Inst, Biomed Om Grp, Cheongju, Chungbuk, South Korea

[3] Univ Bonn, Life & Brain Ctr, Inst Reconstruct Neurobiol, Bonn, Germany

[4] Hertie Fdn, Bonn, Germany

来源：

NATURE PROTOCOLS | 2015年 / 10卷 / 07期

基金：

美国国家卫生研究院; 新加坡国家研究基金会;

关键词：

INTRACELLULAR A-BETA; PROGENITOR CELLS; DIFFERENTIATION; BRAIN; EXPRESSION; CHALLENGES; NUCLEASES; SURVIVAL; NEURONS; GROWTH;

D O I：

10.1038/nprot.2015.065

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Stem 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 3D culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of amyloid-beta (A beta) 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 (hNPCNPCNPCs) with familial AD mutations, the differentiation of the hNPCNPCNPCs in a 3D matrix and the analysis of AD pathogenesis. The 3D culture generation takes 1-2 d. The aggregation of A beta is observed after 6 weeks of differentiation, followed by robust tau pathology after 10-14 weeks.

引用

页码：985 / 1006

页数：22

共 44 条

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