Human Neurospheroid Arrays for In Vitro Studies of Alzheimer's Disease

Sci Rep. 2018 Feb 5;8(1):2450. doi: 10.1038/s41598-018-20436-8.


Neurospheroids are commonly used for in vitro disease modeling and drug screening. However, the heterogeneity in size of the neurospheroids mixtures available through current methods limits their utility when employed for basic mechanistic studies of neurodegenerative diseases or screening for new interventions. Here, we generate neurospheroids from immortalized neural progenitor cells and human induced pluripotent stem cells that are uniform in size, into large-scale arrays. In proof of concept experiments, we validate the neurospheroids array as a sensitive and robust tool for screening compounds over extended time. We show that when suspended in three-dimensional extracellular matrix up to several weeks, the stem cell-derived neurospheroids display extensive neurite outgrowth and extend thick bundles of dendrites outward. We also cultivate genetically-engineered stem cell-derived neurospheroids with familial Alzheimer's disease mutations for eight weeks in our microarray system. Interestingly, we observed robust accumulation of amyloid-β and phosphorylated tau, key hallmarks of Alzheimer's disease. Overall, our in vitro model for engineering neurospheroid arrays is a valuable tool for studying complex neurodegenerative diseases and accelerating drug discovery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alzheimer Disease / genetics
  • Alzheimer Disease / pathology
  • Amyloid beta-Peptides / genetics*
  • Amyloid beta-Peptides / metabolism
  • Biomarkers / metabolism
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Engineering / instrumentation*
  • Cell Engineering / methods
  • Cell Size
  • Gene Expression
  • Genes, Reporter
  • Glutamate Decarboxylase / genetics
  • Glutamate Decarboxylase / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Models, Biological
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / ultrastructure
  • Neuronal Outgrowth
  • Neurons / metabolism
  • Neurons / ultrastructure*
  • Receptors, N-Methyl-D-Aspartate / genetics
  • Receptors, N-Methyl-D-Aspartate / metabolism
  • Spheroids, Cellular / metabolism
  • Spheroids, Cellular / ultrastructure*
  • Tissue Array Analysis / methods*
  • Tyrosine 3-Monooxygenase / genetics
  • Tyrosine 3-Monooxygenase / metabolism
  • tau Proteins / genetics*
  • tau Proteins / metabolism


  • Amyloid beta-Peptides
  • Biomarkers
  • MAP2 protein, human
  • MAPT protein, human
  • Microtubule-Associated Proteins
  • NR2B NMDA receptor
  • Receptors, N-Methyl-D-Aspartate
  • tau Proteins
  • Green Fluorescent Proteins
  • Tyrosine 3-Monooxygenase
  • Glutamate Decarboxylase
  • glutamate decarboxylase 2