Development of microchannel device for long term neuron culture

Abstract

Micro technology holds the promise for advanced cell culture systems which can better mimic the in vivo environment and serve as good disease models. This advantage of micro technology can be leveraged to culture nerve cells in vitro in confined space and nanolitres of culture media. In this work, a tubeless no-flow condition microchannel device is developed with the capabilities to sustain in vitro neural culture for long periods of time. Primary hippocampal neurons from E-18 Sprague-Dawley (SD) rat are cultured inside poly-(dimethylsiloxane) (PDMS) microchannel device assembled on Poly-D-Lysine (PDL) coated substrate. Neurons are cultured inside closed microchannels of variable geometry for over 1 month in serum free media condition. The neuron culture is characterized based on different parameters and effect of channel geometry on neuron development is investigated. Further, this microchannel neuron culture device is integrated with Multielectrode array (MEA) for functional characterization and spontaneous extracellular signals are successfully recorded from neurons inside the channels. This microchannel neuron culture device can help to probe and manipulate the neuron microenvironment and serve as good tool for neuron circuit design, neuron based drug screening assays, neural tissue engineering and in vitro neurodegenerative disease models.