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

384-well plates Cell culture Drug discovery Induced pluripotent stem cells Microplates Neuronal cells

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2019
Historique:
entrez: 25 5 2019
pubmed: 28 5 2019
medline: 13 3 2020
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 31124113
doi: 10.1007/978-1-4939-9477-9_14
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

159-164

Subventions

Organisme : Medical Research Council
ID : MC_EX_G0800785
Pays : United Kingdom

Auteurs

Daniel Little (D)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK. d.little@ucl.ac.uk.

Christin Luft (C)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Oliver Pezzini-Picart (O)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Olukunbi Mosaku (O)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Robin Ketteler (R)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Michael J Devine (MJ)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Paul Gissen (P)

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
Great Ormond Street Institute of Child Health, University College London, London, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, University College London, London, UK.

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Classifications MeSH