Title: Controlled placement of multiple CNS cell populations to create complex neuronal cultures

Abstract

In vitro brain-on-a-chip platforms hold promise in many areas including: drug discovery, evaluating effects of toxicants and pathogens, and disease modelling. A more accurate recapitulation of the intricate organization of the brain in vivo may require a complex in vitro system including organization of multiple neuronal cell types in an anatomically-relevant manner. Most approaches for compartmentalizing or segregating multiple cell types on microfabricated substrates use either permanent physical surface features or chemical surface functionalization. This study describes a removable insert that successfully deposits neurons from different brain areas onto discrete regions of a microelectrode array (MEA) surface, achieving a separation distance of 100 μm. The regional seeding area on the substrate is significantly smaller than current platforms using comparable placement methods. The non-permanent barrier between cell populations allows the cells to remain localized and attach to the substrate while the insert is in place and interact with neighboring regions after removal. The insert was used to simultaneously seed primary rodent hippocampal and cortical neurons onto MEAs. These cells retained their morphology, viability, and function after seeding through the cell insert through 28 days in vitro (DIV). Co-cultures of the two neuron types developed processes and formed integrated networks between themore » different MEA regions. Electrophysiological data demonstrated characteristic bursting features and waveform shapes that were consistent for each neuron type in both mono- and co-culture. Additionally, hippocampal cells co-cultured with cortical neurons showed an increase in withinburst firing rate (p = 0.013) and percent spikes in bursts (p = 0.002), changes that imply communication exists between the two cell types in co-culture. The cell seeding insert described in this work is a simple but effective method of separating distinct neuronal populations on microfabricated devices, and offers a unique approach to developing the types of complex in vitro cellular environments required for anatomically-relevant brain-on-a-chip devices.« less

Authors:

Soscia, D. ;

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Belle, A.; Fischer, N.; Enright, H.; Sales, A.; Osburn, J.; Benett, W.; Mukerjee, E.; Kulp, K.; Pannu, S.; Wheeler, E.; Lytton, ed., William W.

Publication Date:

2017-11-21

Research Org.:

Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

Sponsoring Org.:

USDOE Office of Science (SC)

OSTI Identifier:

1409719

Alternate Identifier(s):

OSTI ID: 1627842

Grant/Contract Number:  

AC52-07NA27344

Resource Type:

Published Article

Journal Name:

PLoS ONE

Additional Journal Information:

Journal Name: PLoS ONE Journal Volume: 12 Journal Issue: 11; Journal ID: ISSN 1932-6203

Publisher:

Public Library of Science

Country of Publication:

United States

Language:

English

Subject:

60 APPLIED LIFE SCIENCES; Hippocampus; Neurons; Action potentials; Electrophysiology; Fluorescence imaging; Cell cultures; Electrode recording; Chemical deposition