Electrophysiological measurements in three-dimensional in vivo-mimetic organotypic cell cultures: Preliminary studies with hen embryo brain spheroids

Uroukov, Ivan S.; Ma, Mingwen; Bull, Larry; Purcell, Wendy M.

Abstract

Using three-dimensional artificial tissue constructs shown to offer organotypic functionality, hen embryo brain spheroids were used as a novel electrophysiological paradigm. For the first time, single spontaneous action potentials were recorded from spheroids in culture at day 7 in vitro (DIV) using multi-electrode arrays. At DIV14 'bursting behaviour' was observed. Simple stimulation was found to induce an increase in spiking frequency with an effect that ramped up over DIV7-14. By DIV14, the frequency under stimulation was typically over twice that of the corresponding spontaneous spiking. These results indicate strong self-organizing processes in vitro within the neuronal networks of the three-dimensional spheroid cell cultures. The organotypic in vivo-mimetic nature of the spheroid paradigm was confirmed by electron microscopy that revealed an outer layer of glial cells, a glial limitans, while immunostaining for Neurofilament and Glial Fibrilliary Acidic Protein demonstrated neuronal cells with a centralized neuronal and synaptic distribution. Basic biochemical functionality was also determined and Acetylcholinesterase measured, indicating the activity of acetylcholine receptors. Thus the organotypic hen embryo brain spheroid model may offer a new paradigm in which to explore neuronal networks. © 2006 Elsevier Ireland Ltd. All rights reserved.

• Journal Article Type: Article
• Publication Date: Aug 14, 2006
• Journal: Neuroscience Letters
• Print ISSN: 0304-3940
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 404
• Issue: 1-2
• Pages: 33-38
• DOI: https://doi.org/10.1016/j.neulet.2006.05.016
• Keywords: brain, cell culture, multi-electrode array, neuronal networks, spike generation
• Public URL: https://uwe-repository.worktribe.com/output/1045554
• Publisher URL: http://dx.doi.org/10.1016/j.neulet.2006.05.016