Long-term adherence of human brain cells in vitro is enhanced by charged amine-based plasma polymer coatings

Bridget Milky, Michael Zabolocki, Sameer A. Al-Bataineh, Mark van den Hurk, Zarina Greenberg, Lucy Turner, Paris Mazzachi, Amber Williams, Imanthi Illeperuma, Robert Adams, Brett W. Stringer, Rebecca Ormsby, Santosh Poonnoose, Louise E. Smith, Marta Krasowska, Jason D. Whittle, Antonio Simula, Cedric Bardy

Research output: Contribution to journalArticlepeer-review

Abstract

Advances in cellular reprogramming have radically increased the use of patient-derived cells for neurological research in vitro. However, adherence of human neurons on tissue cultureware is unreliable over the extended periods required for electrophysiological maturation. Adherence issues are particularly prominent for transferable glass coverslips, hindering imaging and electrophysiological assays. Here, we assessed thin-film plasma polymer treatments, polymeric factors, and extracellular matrix coatings for extending the adherence of human neuronal cultures on glass. We find that positive-charged, amine-based plasma polymers improve the adherence of a range of human brain cells. Diaminopropane (DAP) treatment with laminin-based coating optimally supports long-term maturation of fundamental ion channel properties and synaptic activity of human neurons. As proof of concept, we demonstrated that DAP-treated glass is ideal for live imaging, patch-clamping, and optogenetics. A DAP-treated glass surface reduces the technical variability of human neuronal models and enhances electrophysiological maturation, allowing more reliable discoveries of treatments for neurological and psychiatric disorders.

Original languageEnglish
Pages (from-to)489-506
Number of pages18
JournalStem Cell Reports
Volume17
Issue number3
DOIs
Publication statusPublished - 8 Mar 2022

Bibliographical note

Funding Information:
This work was generously supported by Perpetual Impact Philanthropy, the Brain Foundation, the Rebecca L. Cooper foundation, Australia, the Ian Potter Foundation, Michael and Angelique Boileau Corporate Philanthropy, The Hospital Research Foundation: Parkinson's South Australia, The Michael J Fox Foundation, The Shake It Up Foundation Australia, The Grosset Gaia Foundation, The Mark Hugues Foundation, Cancer Council SA, Cancer Australia, Cooperative Research Centre for Cell Therapy Manufacturing, Australia, and the MRFF Australian Government and Department of Health, Australian Research Council LIEF grant (to C.B.); the Netherlands Organisation for Scientific Research Rubicon Fellowship 019.163LW.032 (to M.v.d.H.); the Flinders University Research Scholarship (to M.Z. and B.M.); and the Australian Government Research Training Program Scholarship (to R.A.). Flow cytometry analysis and cell sorting were performed at the South Australian Health Medical Research Institute (SAHMRI) in the ACRF Cellular Imaging and Cytometry Core Facility. The facility is generously supported by the Detmold Hoopman Group, the Australian Cancer Research Foundation, and the Australian government through the Zero Childhood Cancer Program.

Funding Information:
This work was generously supported by Perpetual Impact Philanthropy, the Brain Foundation , the Rebecca L. Cooper foundation, Australia , the Ian Potter Foundation , Michael and Angelique Boileau Corporate Philanthropy, The Hospital Research Foundation : Parkinson’s South Australia, The Michael J Fox Foundation, The Shake It Up Foundation Australia, The Grosset Gaia Foundation, The Mark Hugues Foundation, Cancer Council SA, Cancer Australia, Cooperative Research Centre for Cell Therapy Manufacturing, Australia, and the MRFF Australian Government and Department of Health , Australian Research Council LIEF grant (to C.B.); the Netherlands Organisation for Scientific Research Rubicon Fellowship 019.163LW.032 (to M.v.d.H.); the Flinders University Research Scholarship (to M.Z. and B.M.); and the Australian Government Research Training Program Scholarship (to R.A.). Flow cytometry analysis and cell sorting were performed at the South Australian Health Medical Research Institute ( SAHMRI ) in the ACRF Cellular Imaging and Cytometry Core Facility. The facility is generously supported by the Detmold Hoopman Group, the Australian Cancer Research Foundation , and the Australian government through the Zero Childhood Cancer Program.

Publisher Copyright:
© 2022 The Authors

Keywords

  • diaminopropane
  • electrophysiology
  • glass coverslips
  • human neurons
  • imaging
  • iPSC
  • neural stem cells
  • plasma polymers
  • pluripotent stem cells
  • tissue cultureware

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