A microfluidic platform for controlled biochemical stimulation of twin neuronal networks |
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| Authors: | Biffi Emilia Piraino Francesco Pedrocchi Alessandra Fiore Gianfranco B Ferrigno Giancarlo Redaelli Alberto Menegon Andrea Rasponi Marco |
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| Institution: | 1Politecnico di Milano, Bioengineering Department, Neuroengineering and Medical Robotics Laboratory, p.zza Leonardo da Vinci 32, 20133 Milano, Italy;2Advanced Light and Electron Microscopy Bio-Imaging Centre, Experimental Imaging Centre, San Raffaele Scientific Institute, via Olgettina 58, 20132 Milano, Italy;3Politecnico di Milano, Bioengineering Department, Biomechanics Group, p.zza Leonardo da Vinci 32, 20133 Milano, Italy |
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| Abstract: | Spatially and temporally resolved delivery of soluble factors is a key feature for pharmacological applications. In this framework, microfluidics coupled to multisite electrophysiology offers great advantages in neuropharmacology and toxicology. In this work, a microfluidic device for biochemical stimulation of neuronal networks was developed. A micro-chamber for cell culturing, previously developed and tested for long term neuronal growth by our group, was provided with a thin wall, which partially divided the cell culture region in two sub-compartments. The device was reversibly coupled to a flat micro electrode array and used to culture primary neurons in the same microenvironment. We demonstrated that the two fluidically connected compartments were able to originate two parallel neuronal networks with similar electrophysiological activity but functionally independent. Furthermore, the device allowed to connect the outlet port to a syringe pump and to transform the static culture chamber in a perfused one. At 14 days invitro, sub-networks were independently stimulated with a test molecule, tetrodotoxin, a neurotoxin known to block action potentials, by means of continuous delivery. Electrical activity recordings proved the ability of the device configuration to selectively stimulate each neuronal network individually. The proposed microfluidic approach represents an innovative methodology to perform biological, pharmacological, and electrophysiological experiments on neuronal networks. Indeed, it allows for controlled delivery of substances to cells, and it overcomes the limitations due to standard drug stimulation techniques. Finally, the twin network configuration reduces biological variability, which has important outcomes on pharmacological and drug screening. |
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