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.
|Original language||English (US)|
|Number of pages||8|
|State||Published - 2008|
- Cortical neurons
ASJC Scopus subject areas
- Biomedical Engineering
- Electrical and Electronic Engineering