A compartmentalized culture device for studying the axons of CNS neurons
| Autor: | Wen-Cheng Yang, Hui-Wen Chung, Chien-Chung Fu, Yun-Hsin Hsu, Mei-Yun Cheng, Chih-Fan Chuang, Tsung-Kai Huang, Hui-Yu Chen, Yen-Chung Chang, Hsing-Hua Ho, Wan-Chong Leong |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Paclitaxel Neurite Central nervous system Cell Culture Techniques Biophysics Glutamic Acid Hippocampal formation Biology Microtubules Biochemistry 03 medical and health sciences 0302 clinical medicine Microtubule medicine Animals Compartment (development) Dimethylpolysiloxanes Molecular Biology Cells Cultured Neurons Dendrites Cell Biology Anatomy Microfluidic Analytical Techniques Embryo Mammalian Axons Rats 030104 developmental biology medicine.anatomical_structure Membrane Microscopy Fluorescence nervous system Cell culture Neuron 030217 neurology & neurosurgery |
| Zdroj: | Analytical Biochemistry. 539:11-21 |
| ISSN: | 0003-2697 |
| Popis: | We report here the development of a compartmentalized culture device that allows the spatial separation of the somatodendrites and axons of central nervous system (CNS) neurons. The device consists of two compartments separated by a septum constructed by attaching a porous polycarbonate track etch (PCTE) filter on top of a microchannel-filled polydimethylsiloxane (PDMS) membrane. The surface and microchannels of the septum are coated and filled, respectively, with materials that support neuron growth and neurite migration. When rat hippocampal neurons are cultured in the top compartment, axons are the only processes that can migrate through the septum to the bottom compartment. The axons in the bottom compartment can be studied directly in real-time or through immunofluorescence staining after fixation. Axons containing ∼3 μg protein can be isolated from each device for biochemical analyses. In addition, the septum also impedes the movement of small molecules between the top and bottom compartments. This feature allows the somatodendrites and axons of neurons, which occupy the top and bottom compartments of the device, respectively, to be manipulated independently. The potential applications of the device as a tool in diverse studies concerning neuronal axons and in screening reagents that regulate axonal functions have also been discussed. |
| Databáze: | OpenAIRE |
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