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

Q3 Biochemistry, Genetics and Molecular Biology

Current Protocols in Cell Biology Pub Date : 2019-10-04 DOI:10.1002/cpcb.98

Aurelie de Rus Jacquet

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引用次数: 14

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

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多能干细胞多巴胺能神经元与星形胶质细胞的制备与共培养

基于诱导多能干细胞(iPSC)的模型是研究帕金森病等神经退行性疾病的有力工具。患者来源的神经元和星形胶质细胞的分化使研究疾病发生和发展的分子机制成为可能。特别是，这两种细胞类型可以单独或共同培养，以研究细胞自主和非细胞自主贡献者对神经退行性疾病的影响。我们开发了一种简化的程序来生产高质量/高纯度的多巴胺能神经元和星形胶质细胞的培养，它们来自同一群中脑底板祖细胞。本单元描述了差异化，质量控制，培养参数和故障排除提示，以确保研究结果的最高质量和可重复性。©2019作者。基本方案1:多能干细胞分化为中脑模式神经祖细胞支持方案:神经祖细胞质量控制基本方案2:神经祖细胞分化为星形胶质细胞基本方案3:神经祖细胞分化为多巴胺能神经元基本方案4:多能干细胞衍生的神经元和星形胶质细胞共培养

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Current Protocols in Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology

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期刊介绍： Developed by leading scientists in the field, Current Protocols in Cell Biology is an essential reference for researchers who study the relationship between specific molecules and genes and their location, function and structure at the cellular level. Updated every three months in all formats, CPCB is constantly evolving to keep pace with the very latest discoveries and developments.