Isolation of Human CD49f+ Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays

STAR Protoc. 2020 Nov 10;1(3):100172. doi: 10.1016/j.xpro.2020.100172. eCollection 2020 Dec 18.

Abstract

Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem cell (iPSC)-based cultures, neural organoids, and primary tissue, using the surface marker CD49f. Moreover, we provide protocols for in vitro co-cultures of human iPSC-derived neurons and astrocytes, as well as for neurotoxicity assays that expose neurons to conditioned media from reactive astrocytes. For complete details on the use and execution of this protocol, please refer to Barbar et al. (2020).

Keywords: Cell Biology; Neuroscience; Stem Cells.

Publication types

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

MeSH terms

  • Astrocytes / cytology
  • Astrocytes / drug effects
  • Astrocytes / metabolism*
  • Biological Assay / methods*
  • Cell Differentiation / drug effects
  • Cell Separation*
  • Cells, Cultured
  • Coculture Techniques
  • Culture Media, Conditioned / pharmacology
  • Flow Cytometry
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism*
  • Integrin alpha6 / metabolism*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurotoxins / toxicity*
  • Toxicity Tests*

Substances

  • Culture Media, Conditioned
  • Integrin alpha6
  • Neurotoxins