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

  • Daniel LittleEmail author
  • Christin Luft
  • Oliver Pezzini-Picart
  • Olukunbi Mosaku
  • Robin Ketteler
  • Michael J. Devine
  • Paul Gissen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1994)

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.

Key words

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

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Daniel Little
    • 1
    Email author
  • Christin Luft
    • 1
  • Oliver Pezzini-Picart
    • 1
  • Olukunbi Mosaku
    • 1
  • Robin Ketteler
    • 1
  • Michael J. Devine
    • 1
  • Paul Gissen
    • 2
    • 3
    • 4
  1. 1.MRC Laboratory for Molecular Cell BiologyUniversity College LondonLondonUK
  2. 2.MRC Laboratory for Molecular Cell BiologyUniversity College LondonLondonUK
  3. 3.Great Ormond Street Institute of Child HealthUniversity College LondonLondonUK
  4. 4.NIHR Great Ormond Street Hospital Biomedical Research CentreUniversity College LondonLondonUK

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