Microfluidic Multicompartment Device for Neuroscience Research
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- 作者：Anne M. Taylor ; Seog Woo Rhee ; Christina H. Tu ; David H. Cribbs ; Carl W. Cotman ; and Noo Li Jeon
- 刊名：Langmuir
- 出版年：2003
- 出版时间：March 4, 2003
- 年：2003
- 卷：19
- 期：5
- 页码：1551 - 1556
- 全文大小：787K
- 年卷期：v.19,no.5(March 4, 2003)
- ISSN：1520-5827
文摘
This paper describes and characterizes a novel microfabricated neuronal culture device. This devicecombines 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 PDMSdevice is placed on a tissue culture dish (polystyrene) or glass substrate, forming two compartments withvolumes of less than 2 
L each. These two compartments are separated by a physical barrier in which anumber of micron-size grooves are embedded to allow growth of neurites across the compartments whilemaintaining 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 isbetween 50 and 70% after 7 days in culture; this is slightly lower than but comparable to values for a controlgrown 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 themicrogrooves for fluid transport, insults are contained in the neuritic compartment without appreciableleakage into the somal compartment for over 15 h. Finally, we demonstrate the use of polylysine patterningin 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 micropatterningtechniques, combined with fluidically isolated compartments within the culture area, offers significantadvantages over standard open culture methods and other conventional methods for manipulating distinctneuronal microenvironments.
L each. These two compartments are separated by a physical barrier in which anumber of micron-size grooves are embedded to allow growth of neurites across the compartments whilemaintaining 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 isbetween 50 and 70% after 7 days in culture; this is slightly lower than but comparable to values for a controlgrown 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 themicrogrooves for fluid transport, insults are contained in the neuritic compartment without appreciableleakage into the somal compartment for over 15 h. Finally, we demonstrate the use of polylysine patterningin 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 micropatterningtechniques, combined with fluidically isolated compartments within the culture area, offers significantadvantages over standard open culture methods and other conventional methods for manipulating distinctneuronal microenvironments.