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

Abstract

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 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.

Microglial dysfunction as a key pathological change in adrenomyeloneuropathy

“…Immortalized hNPC cell line ReNcell VM (ReN) was used to differentiate into neurons per published protocol 16. Simply, ReN cells were grown in proliferation medium (DMEM/F12; Gibco/Life Technologies, Grand Island, NY) with 2 µg/ml of heparin (stock; STEMCELL Technologies, Cambridge, MA), 2% B27 (Life Technologies, Carlsbad, CA), 20ng/ml of epidermal growth factor (EGF), 20ng/ml of basic fibroblast growth factor, and 1% of 100 × penicillin/streptomycin (Gibco/Life Technologies) until confluency.…”

Developments in 3D neural cell culture models: the future of neurotherapeutics testing?

“…Mammary gland cells were observed to have differing morphologies and behaviors when grown in Matrigel versus type I collagen [24]. This effect does not appear to be present in growing neural cells, as investigators have used both Matrigel and type I collagen in separate scaffold-based cell culture studies without mention of significant anomalies [3,22]. However, there has yet to be any thorough comparison across scaffold-based models, which may vary considerably by scaffold fabrication method and materials.…”

“…Synthetic materials such as polycaprolactone, polyethylene glycol, and even polystyrene have been used as scaffolds for cell adherence and support [5,19,20]. Another method for scaffold-based neural cell culture provides cells with natural ECM proteins such as laminin or collagen [3,21,22]. Type I collagen with hyaluronic acid and alginate with laminin are some examples of natural material combinations used as cell scaffolds [3,21].…”

[P2–068]: SOLUBLE AMYLOID‐BETA AGGREGATES FROM HUMAN ALZHEIMER's DISEASE BRAINS

“…Thus, characterizing a range of animal models of AD-like pathology is a top priority. It is possible, however, that none of the existing animal models accurately recapitulate human soluble Aβ aggregates, and that new model systems will need to be developed 55 . Furthermore, the purification scheme developed here may be thought of as a generalizable framework for assessment of proteins aggregates relevant to human neurodegenerative diseases.…”