Little Daniel, Luft Christin, Pezzini-Picart Oliver, Mosaku Olukunbi, Ketteler Robin, Devine Michael J, Gissen Paul
MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
Great Ormond Street Institute of Child Health, University College London, London, UK.
Methods Mol Biol. 2019;1994:159-164. doi: 10.1007/978-1-4939-9477-9_14.
Induced pluripotent stem cell (iPSC) derived neurons are an excellent in vitro model of neurological diseases that are often used in early stage drug discovery projects. Thus far, the use of iPSC-derived cells in small molecule drug screening has been limited, and one of the reasons for this has been the challenge of miniaturization of iPSC culture and differentiation in low volume microwell plate formats. Here we describe a method of seeding iPSC-derived neurons into 384-well plates towards the end of the differentiation procedure. This method covers coating the plates with substrates to aid attachment, dissociation of the cells into a single cell suspension, and seeding onto 384-well plates to give an even distribution of neurons. This method facilitates the use of iPSC-derived neurons for high-content imaging, whole-well assays, and small-molecule drug screening.
诱导多能干细胞（iPSC）衍生的神经元是神经疾病的一种优秀体外模型，常用于早期药物发现项目。到目前为止，iPSC衍生细胞在小分子药物筛选中的应用一直受到限制，其中一个原因是在低体积微孔板格式中iPSC培养和分化的小型化面临挑战。在此，我们描述了一种在分化过程接近尾声时将iPSC衍生的神经元接种到384孔板中的方法。该方法包括用底物包被板以促进细胞附着，将细胞解离成单细胞悬液，并接种到384孔板上以使神经元均匀分布。此方法有助于将iPSC衍生的神经元用于高内涵成像、全孔分析和小分子药物筛选。
