Microfluidic Multicompartment Device for Neuroscience Research

Abstract

This paper describes and characterizes a novel microfabricated neuronal culture device. This device combines microfabrication, microfluidic, and surface micropatterning techniques to create a multicompartment 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 of less than 2 μL 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 and 70% after 7 days in culture; this is slightly lower than but comparable to values for a control 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 microgrooves for fluid transport, insults are contained in the neuritic compartment without appreciable leakage into the somal compartment for over 15 h. 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, offers significant advantages over standard open culture methods and other conventional methods for manipulating distinct neuronal microenvironments.

Patterned cell culture inside microfluidic devices

“…A compartmented microfluidic neuronal culture device was fabricated in PDMS to achieve fluidically isolated microenvironments for somas and neurites. 16 Fig. 4a shows the schematic drawing of the device.…”

“…The microfluidic cell culture device was fabricated in PDMS using rapid prototyping and soft lithography according to published procedures. 1,16,17 The master for the neuronal culture device was fabricated by patterning two layers of photoresist. 16 Briefly, first layer of photoresist, 3 mm thick was obtained by spinning SU-8 5 (Microchem, Newton, MA) negative photoresist at 3,500 rpm for 60 s. A 20,000 dpi high-resolution printer (CAD/Art Services, San Diego, CA) was used to generate the first transparency mask to create the microchannels (10 mm wide and spaced 50 mm).…”

A microfluidic culture platform for CNS axonal injury, regeneration and transport

“…The high fluidic resistance of the microgrooves produces a small but sustained flow between the compartments that counteracts diffusion 23,24 . To demonstrate fluidic integrity of the compartments, we incubated Texas red dextran (3,000 Da; 20 μM) in the axonal compartment for over 20 h. We detected no trace of fluorescence within the somal compartment by either UV spectroscopy (equivalent to background) or confocal microscopy (Fig.…”

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

“…Recently, Taylor et al 195 created a neuron culture PDMS device with the capability to direct neuronal attachment and neurite outgrowth and to fluidically isolate compartments within the culture area. Essentially, the device consisted of two chambers interconnected with a set of 120 parallel PDMS microchannels 10 μm in width, 3 μm in height, and 150 μm in length.…”