Neuronal-glial populations form functional networks in a biocompatible 3D scaffold.
Monolayers of neurons and glia have been employed for decades as tools for the study of cellular physiology and as the basis for a variety of standard toxicological assays. A variety of three dimensional (3D) culture techniques have been developed with the aim to produce cultures that recapitulate desirable features of intact. In this study, we investigated the effect of preparing primary mouse mixed neuron and glial cultures in the inert 3D scaffold, Alvetex. Using planar multielectrode arrays, we compared the spontaneous bioelectrical activity exhibited by neuroglial networks grown in the scaffold with that seen in the same cells prepared as conventional monolayer cultures. Two dimensional (monolayer; 2D) cultures exhibited a significantly higher spike firing rate than that seen in 3D cultures although no difference was seen in total signal power (<50Hz) while pharmacological responsiveness of each culture type to antagonism of GABAAR, NMDAR and AMPAR was highly comparable. Interestingly, correlation of burst events, spike firing and total signal power (<50Hz) revealed that local field potential events were associated with action potential driven bursts as was the case for 2D cultures. Moreover, glial morphology was more physiologically normal in 3D cultures. These results show that 3D culture in inert scaffolds represents a more physiologically normal preparation which has advantages for physiological, pharmacological, toxicological and drug development studies, particularly given the extensive use of such preparations in high throughput and high content systems.
School of Chemistry, Food & Nutritional Sciences and Pharmacy, University of Reading, Whiteknights, Reading, Berkshire RG6 6AP, UK; Portsmouth Brain Tumour Research Centre, University of Portsmouth, White Swan Road, Portsmouth PO1 2DT, UK. Electronic address: immy.smith@port.ac.uk.
Imogen
I
Smith
School of Chemistry, Food & Nutritional Sciences and Pharmacy, University of Reading, Whiteknights, Reading, Berkshire RG6 6AP, UK. Electronic address: m.haag@pgr.reading.ac.uk.
Marcus
M
Haag
School of Chemistry, Food & Nutritional Sciences and Pharmacy, University of Reading, Whiteknights, Reading, Berkshire RG6 6AP, UK; School of Pharmacy, University of Bradford, Bradford, West Yorkshire BD7 1DP, UK. Electronic address: c.ugbode@bradford.ac.uk.
Christopher
C
Ugbode
School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, UK. Electronic address: daniel.tams@durham.ac.uk.
Daniel
D
Tams
School of Pharmacy, University of Bradford, Bradford, West Yorkshire BD7 1DP, UK. Electronic address: m.rattray@bradford.ac.uk.
Marcus
M
Rattray
School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, UK. Electronic address: stefan.przyborski@durham.ac.uk.
Stefan
S
Przyborski
School of Chemistry, Food & Nutritional Sciences and Pharmacy, University of Reading, Whiteknights, Reading, Berkshire RG6 6AP, UK. Electronic address: a.bithell@reading.ac.uk.
Angela
A
Bithell
School of Chemistry, Food & Nutritional Sciences and Pharmacy, University of Reading, Whiteknights, Reading, Berkshire RG6 6AP, UK. Electronic address: b.j.whalley@reading.ac.uk.
Benjamin J
BJ
Whalley
D000200:Action Potentials; D000818:Animals; D051379:Mice; D009457:Neuroglia / Q000166:cytology* / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D046509:Tissue Culture Techniques / Q000379:methods*; D054457:Tissue Scaffolds
3D Neuronal culture; Biocompatible scaffold; Cortical cultures; Microelectrode array; Murine