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Microfluidic culture platform for neuroscience research

Nature Protocols volume 1, pages 2128–2136 (2006)Cite this article

Abstract

This protocol describes the fabrication and use of a microfluidic device to culture central nervous system (CNS) and peripheral nervous system neurons for neuroscience applications. This method uses replica-molded transparent polymer parts to create miniature multi-compartment cell culture platforms. The compartments are made of tiny channels with dimensions of tens to hundreds of micrometers that are large enough to culture a few thousand cells in well-controlled microenvironments. The compartments for axon and somata are separated by a physical partition that has a number of embedded micrometer-sized grooves. After 3–4 days in vitro (DIV), cells that are plated into the somal compartment have axons that extend across the barrier through the microgrooves. The culture platform is compatible with microscopy methods such as phase contrast, differential interference microscopy, fluorescence and confocal microscopy. Cells can be cultured for 2–3 weeks within the device, after which they can be fixed and stained for immunocytochemistry. Axonal and somal compartments can be maintained fluidically isolated from each other by using a small hydrostatic pressure difference; this feature can be used to localize soluble insults to one compartment for up to 20 h after each medium change. Fluidic isolation enables collection of pure axonal fraction and biochemical analysis by PCR. The microfluidic device provides a highly adaptable platform for neuroscience research and may find applications in modeling CNS injury and neurodegeneration. This protocol can be completed in 1–2 days.

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Figure 1: Schematic of master mold fabrication.
Figure 2: Schematic of PDMS device fabrication by soft lithography.
Figure 3: Two modes of assembling devices.
Figure 4: Confocal micrograph of Tau (green) and MAP5 (red) immunostained neurons (a) and phase micrographs of 6 DIV neurons (b,c) cultured in the microfluidic device.
Figure 5: Schematic of axotomy procedure in the microfluidic device.
Figure 6
Figure 7: RNA blot of samples isolated from somal and axonal chambers.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, Irvine, 3410

    Jeong Won Park, Behrad Vahidi, Anne M Taylor & Noo Li Jeon

  2. Natural Science II, Irvine, 92697, California, USA

    Jeong Won Park, Behrad Vahidi, Anne M Taylor & Noo Li Jeon

  3. Department of Chemistry, Kongju National University, 182 Shinkwan-dong, Kongju, 314-701, Chungnam, Republic of Korea

    Seog Woo Rhee

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  1. Jeong Won Park
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Correspondence to Noo Li Jeon.

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Three of the authors (AMT, SWR, and NLJ) have personal financial interests associated with patent applications.

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Park, J., Vahidi, B., Taylor, A. et al. Microfluidic culture platform for neuroscience research. Nat Protoc 1, 2128–2136 (2006). https://doi.org/10.1038/nprot.2006.316

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