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

doi:10.1016/j.stemcr.2022.01.013

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

College of Medicine and Public Health

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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 language	English
Pages (from-to)	489-506
Number of pages	18
Journal	Stem Cell Reports
Volume	17
Issue number	3
DOIs	https://doi.org/10.1016/j.stemcr.2022.01.013
Publication status	Published - 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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Milky, B., Zabolocki, M., Al-Bataineh, S. A., van den Hurk, M., Greenberg, Z., Turner, L., Mazzachi, P., Williams, A., Illeperuma, I., Adams, R., Stringer, B. W., Ormsby, R., Poonnoose, S., Smith, L. E., Krasowska, M., Whittle, J. D., Simula, A., & Bardy, C. (2022). Long-term adherence of human brain cells in vitro is enhanced by charged amine-based plasma polymer coatings. Stem Cell Reports, 17(3), 489-506. https://doi.org/10.1016/j.stemcr.2022.01.013

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

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.",

keywords = "diaminopropane, electrophysiology, glass coverslips, human neurons, imaging, iPSC, neural stem cells, plasma polymers, pluripotent stem cells, tissue cultureware",

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

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{\textquoteright}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: {\textcopyright} 2022 The Authors",

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doi = "10.1016/j.stemcr.2022.01.013",

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journal = "Stem Cell Reports",

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Milky, B, Zabolocki, M, Al-Bataineh, SA, van den Hurk, M, Greenberg, Z, Turner, L, Mazzachi, P, Williams, A, Illeperuma, I, Adams, R, Stringer, BW, Ormsby, R , Poonnoose, S, Smith, LE, Krasowska, M, Whittle, JD, Simula, A & Bardy, C 2022, 'Long-term adherence of human brain cells in vitro is enhanced by charged amine-based plasma polymer coatings', Stem Cell Reports, vol. 17, no. 3, pp. 489-506. https://doi.org/10.1016/j.stemcr.2022.01.013

TY - JOUR

T1 - Long-term adherence of human brain cells in vitro is enhanced by charged amine-based plasma polymer coatings

AU - Milky, Bridget

AU - Zabolocki, Michael

AU - Al-Bataineh, Sameer A.

AU - van den Hurk, Mark

AU - Greenberg, Zarina

AU - Turner, Lucy

AU - Mazzachi, Paris

AU - Williams, Amber

AU - Illeperuma, Imanthi

AU - Adams, Robert

AU - Stringer, Brett W.

AU - Ormsby, Rebecca

AU - Poonnoose, Santosh

AU - Smith, Louise E.

AU - Krasowska, Marta

AU - Whittle, Jason D.

AU - Simula, Antonio

AU - Bardy, Cedric

N1 - 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

PY - 2022/3/8

Y1 - 2022/3/8

N2 - 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.

AB - 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.

KW - diaminopropane

KW - electrophysiology

KW - glass coverslips

KW - human neurons

KW - imaging

KW - iPSC

KW - neural stem cells

KW - plasma polymers

KW - pluripotent stem cells

KW - tissue cultureware

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U2 - 10.1016/j.stemcr.2022.01.013

DO - 10.1016/j.stemcr.2022.01.013

M3 - Article

C2 - 35180396

AN - SCOPUS:85125627976

SN - 2213-6711

VL - 17

SP - 489

EP - 506

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 3

ER -

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

Abstract

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Keywords

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