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Systematic Three-Dimensional Coculture Rapidly Recapitulates Interactions between Human Neurons and Astrocytes.

  • Author(s): Krencik, Robert
  • Seo, Kyounghee
  • van Asperen, Jessy V
  • Basu, Nupur
  • Cvetkovic, Caroline
  • Barlas, Saba
  • Chen, Robert
  • Ludwig, Connor
  • Wang, Chao
  • Ward, Michael E
  • Gan, Li
  • Horner, Philip J
  • Rowitch, David H
  • Ullian, Erik M
  • et al.
Abstract

Human astrocytes network with neurons in dynamic ways that are still poorly defined. Our ability to model this relationship is hampered by the lack of relevant and convenient tools to recapitulate this complex interaction. To address this barrier, we have devised efficient coculture systems utilizing 3D organoid-like spheres, termed asteroids, containing pre-differentiated human pluripotent stem cell (hPSC)-derived astrocytes (hAstros) combined with neurons generated from hPSC-derived neural stem cells (hNeurons) or directly induced via Neurogenin 2 overexpression (iNeurons). Our systematic methods rapidly produce structurally complex hAstros and synapses in high-density coculture with iNeurons in precise numbers, allowing for improved studies of neural circuit function, disease modeling, and drug screening. We conclude that these bioengineered neural circuit model systems are reliable and scalable tools to accurately study aspects of human astrocyte-neuron functional properties while being easily accessible for cell-type-specific manipulations and observations.

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