| Info. | Vol.2 - No.1 (2008.03.20) |
|---|---|
| Title | Quantitative Analysis of CNS Axon Regeneration Using a Microfluidic Neuron Culture Device |
| Authors | Jeong Won Park1,3,†, Behrad Vahidi1,†, Hyung Joon Kim1, Seog Woo Rhee2&Noo Li Jeon1 |
| Institutions | 1Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697 2Department of Chemistry, Kongju National University, Chungnam 361-701, Korea 3Department of Life Science, Gwangju Institute of Science and Technonolgy, Gwangju 500-715, Korea †These authors contributed equally to the work Correspondence and requests for materials should be addressed to N.L. Jeon (nljeon@uci.edu ) |
| Abstract | CNS axonal regeneration has been widely studied in order to develop strategies for overcoming Myelin mediated inhibition. However, there are no suitable in vitro methods to distinguish and assess regeneration of severed CNS axons. In this paper we describe the use of a new microfluidic neuron culture device that can be applied quantitatively to investigate the effect of myelin inhibitors on the regeneration of injured CNS axons. The device has two isolated compartments separated by a physical barrier with embedded microgrooves. The device has been successfully used for long-term culture of primary CNS neurons while isolating the somata in one chamber and directing growth of axons to the other chamber. We cultured cortical neurons in the devices and subjected them to reproducible axotomy by vacuum aspiration. After 24 hours, the lengths of regenerated axons were measured for quantitative analysis. NOGO-66 concentrations above 10 nM consistently resulted in ~20% reduction in length of regenerated axons. MAG protein also inhibited axonal regeneration. The length of regenerated axons decreased with addition of myelin inhibitory proteins after injury. These results suggest that the device can serve as an in vitro model for axonal injury and regeneration by simulating the microenvironment around the injury site. |
| Keyword | Cortical neurons, Nogo, MAG, Microfluidics |
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