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BioengineeringbioRxiv

Growing neuronal islands on multi-electrode arrays using an Accurate Positioning-μCP device

Robert Samhaber, Manuel Schottdorf, Ahmed El Hady, Kai Broeking, Andreas Daus, Christiane Thielemann, Walter Stuehmer, Fred Wolf

Abstract

BackgroundMulti-electrode arrays (MEAs) allow non-invasive multiunit recording in-vitro from cultured neuronal networks. For sufficient neuronal growth and adhesion on such MEAs, substrate preparation is required. Plating of dissociated neurons on a uniformly prepared MEAs surface results in the formation of spatially extended random networks with substantial inter-sample variability. Such cultures are not optimally suited to study the relationship between defined structure and dynamics in neuronal networks. To overcome these shortcomings, neurons can be cultured with pre-defined topology by spatially structured surface modification. Spatially structuring a MEA surface accurately and reproducibly with the equipment of a typical cell-culture laboratory is challenging.\n\nNew MethodIn this paper, we present a novel approach utilizing microcontact printing (CP) combined with a custom-made device to accurately position patterns on MEAs with high precision. We call this technique AP-CP (accurate positioning micro-contact printing).\n\nComparison with existing MethodsOther approaches presented in the literature using CP for patterning either relied on facilities or techniques not readily available in a standard cell culture laboratory, or they did not specify means of precise pattern positioning.\n\nConclusionHere we present a relatively simple device for reproducible and precise patterning in a standard cell-culture laboratory setting. The patterned neuronal islands on MEAs provide a basis for high throughput electrophysiology to study the dynamics of single neurons and neuronal networks.

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