Preparation and Co-Culture of iPSC-Derived Dopaminergic Neurons and Astrocytes

Curr Protoc Cell Biol. 2019 Dec;85(1):e98. doi: 10.1002/cpcb.98.

Abstract

Induced pluripotent stem cell (iPSC)-based models are powerful tools to study neurodegenerative diseases such as Parkinson's disease. The differentiation of patient-derived neurons and astrocytes allows investigation of the molecular mechanisms responsible for disease onset and development. In particular, these two cell types can be mono- or co-cultured to study the influence of cell-autonomous and non-cell-autonomous contributors to neurodegenerative diseases. We developed a streamlined procedure to produce high-quality/high-purity cultures of dopaminergic neurons and astrocytes that originate from the same population of midbrain floor-plate progenitors. This unit describes differentiation, quality control, culture parameters, and troubleshooting tips to ensure the highest quality and reproducibility of research results. © 2019 The Authors. Basic Protocol 1: Differentiation of iPSCs into midbrain-patterned neural progenitor cells Support Protocol: Quality control of neural progenitor cells Basic Protocol 2: Differentiation of neural progenitor cells into astrocytes Basic Protocol 3: Differentiation of neural progenitor cells into dopaminergic neurons Basic Protocol 4: Co-culture of iPSC-derived neurons and astrocytes.

Keywords: Parkinson's disease; astrocytes; co-culture; dopaminergic neurons; iPSC differentiation; non-cell-autonomous.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Astrocytes / physiology*
  • Cell Culture Techniques
  • Cell Differentiation
  • Cells, Cultured
  • Coculture Techniques
  • Dopaminergic Neurons / physiology*
  • Humans
  • Induced Pluripotent Stem Cells / physiology*
  • Neurodegenerative Diseases / pathology*
  • Parkinson Disease / pathology*
  • Quality Control