Characterization of neuronal viability and network activity under microfluidic flow.
Drug delivery
Flow
Microculture
Microfluidics
Neuronal culture
Rapid flow
Volume transmission
Journal
Journal of neuroscience methods
ISSN: 1872-678X
Titre abrégé: J Neurosci Methods
Pays: Netherlands
ID NLM: 7905558
Informations de publication
Date de publication:
01 07 2021
01 07 2021
Historique:
received:
02
11
2020
revised:
06
04
2021
accepted:
22
04
2021
pubmed:
2
5
2021
medline:
1
7
2021
entrez:
1
5
2021
Statut:
ppublish
Résumé
Microfluidics technology has the potential to allow precise control of the temporal and spatial aspects of solute concentration, making it highly relevant for the study of volume transmission mechanisms in neural tissue. However, full utilization of this technology depends on understanding how microfluidic flow at the rates needed for rapid solution exchange affects neuronal viability and network activity. We designed a tape-based pressurized microfluidic flow system that is simple to fabricate and can be attached to commercial microelectrode arrays. The device is multi-layered, allowing the inclusion of a porous polycarbonate membrane to isolate neuronal cultures from shear forces while maintaining diffusive exchange of solutes. We used this system to investigate how flow affected survival and spiking patterns of cultured hippocampal neurons. Viability and network activity of the cultures were reduced in proportion to flow rate. However, shear reduction measures did not improve survival or spiking activity; media conditioning in conjunction with culture age proved to be the critical factors for network stability. Diffusion simulations indicate that dilution of a small molecule accounts for the deleterious effects of flow on neuronal cultures. This work establishes the experimental conditions for real time measurement of network activity during rapid solution exchange, using multi-layered chambers with reversible bonding that allow for reuse of microelectrode arrays. With correct media conditioning, the microfluidic flow system allows drug delivery on a subsecond timescale without disruption of network activity or viability, enabling in vitro reproduction of volume transmission mechanisms.
Sections du résumé
BACKGROUND
Microfluidics technology has the potential to allow precise control of the temporal and spatial aspects of solute concentration, making it highly relevant for the study of volume transmission mechanisms in neural tissue. However, full utilization of this technology depends on understanding how microfluidic flow at the rates needed for rapid solution exchange affects neuronal viability and network activity.
NEW METHOD
We designed a tape-based pressurized microfluidic flow system that is simple to fabricate and can be attached to commercial microelectrode arrays. The device is multi-layered, allowing the inclusion of a porous polycarbonate membrane to isolate neuronal cultures from shear forces while maintaining diffusive exchange of solutes. We used this system to investigate how flow affected survival and spiking patterns of cultured hippocampal neurons.
RESULTS
Viability and network activity of the cultures were reduced in proportion to flow rate. However, shear reduction measures did not improve survival or spiking activity; media conditioning in conjunction with culture age proved to be the critical factors for network stability. Diffusion simulations indicate that dilution of a small molecule accounts for the deleterious effects of flow on neuronal cultures.
COMPARISON WITH EXISTING METHODS
This work establishes the experimental conditions for real time measurement of network activity during rapid solution exchange, using multi-layered chambers with reversible bonding that allow for reuse of microelectrode arrays.
CONCLUSIONS
With correct media conditioning, the microfluidic flow system allows drug delivery on a subsecond timescale without disruption of network activity or viability, enabling in vitro reproduction of volume transmission mechanisms.
Identifiants
pubmed: 33932456
pii: S0165-0270(21)00135-7
doi: 10.1016/j.jneumeth.2021.109200
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
109200Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.