Methods Mol Biol. 2019;1994:159-164. doi: 10.1007/978-1-4939-9477-9_14.

Seeding Induced Pluripotent Stem Cell-Derived Neurons onto 384-Well Plates.

Little D¹, Luft C², Pezzini-Picart O², Mosaku O², Ketteler R², Devine MJ², Gissen P^2,3,4.

Abstract

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.