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

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.