Microfluidic Multi-compartment Device for Neuroscience Research
A.M. Taylor, S.W. Rhee, C.H. Tu, D.H.Cribbs, C.W. Cotman and N.L. Jeon
Univeristy of California at Irvine, US
Keywords: microfluidic, multi-compartment device, neuroscience
This paper describes and characterizes a novel microfabricated neuronal culture device. This device combines microfabrication, microfluidic, and surface micropatterning techniques to create a multi-compartment neuronal culturing device that can be used in a number of neuroscience research applications. The device is fabricated in poly(dimethylsiloxane), PDMS, using soft lithography techniques. The PDMS device is placed on a tissue culture dish (polystyrene) or glass substrate, forming two compartments with volumes less than 2 ml each. These two compartments are separated by a physical barrier in which a number of micron-size grooves are embedded to allow growth of neurites across the compartments while maintaining fluidic isolation. Cells are plated into the somal (cell body) compartment and after 3-4 days, neurites extend into the neuritic compartment via the grooves. Viability of the neurons in the devices is between 50-70% after 7 days in culture; this is slightly lower than controls grown on tissue culture dishes. Healthy neuron morphology is evident in both the devices and controls. We demonstrate the ability to use hydrostatic pressure to isolate insults to one compartment and, thus, expose localized areas of neurons to insults applied in soluble form. Due to the high resistance of the micro-grooves for fluid transport, insults are contained in the neuritic compartment without leakage into the somal compartment for over 15 hours. Finally, we demonstrate the use of polylysine patterning in combination with the microfabricated device to facilitate identification and visualization of neurons. The ability to direct sites of neuronal attachment and orientation of neurite outgrowth by micropatterning techniques, combined with fluidically isolated compartments within the culture area offer significant advantages over standard open culture methods and other conventional methods for manipulating distinct neuronal microenvironments.
NSTI Nanotech 2003 Conference Technical Program Abstract