Axon Myelination and Electrical Stimulation in a Microfluidic, Compartmentalized Cell Culture Platform
- 942 Downloads
Axon demyelination contributes to the loss of sensory and motor function following injury or disease in the central nervous system. Numerous reports have demonstrated that myelination can be achieved in neuron/oligodendrocyte co-cultures. However, the ability to selectively treat neuron or oligodendrocyte (OL) cell bodies in co-cultures improves the value of these systems when designing mechanism-based therapeutics. We have developed a microfluidic-based compartmentalized culture system to achieve segregation of neuron and OL cell bodies while simultaneously allowing the formation of myelin sheaths. Our microfluidic platform allows for a high replicate number, minimal leakage, and high flexibility. Using a custom built lid, fit with platinum electrodes for electrical stimulation (10-Hz pulses at a constant 3 V with ~190 kΩ impedance), we employed the microfluidic platform to achieve activity-dependent myelin segment formation. Electrical stimulation of dorsal root ganglia resulted in a fivefold increase in the number of myelinated segments/mm2 when compared to unstimulated controls (19.6 ± 3.0 vs. 3.6 ± 2.3 MBP+ segments/mm2). This work describes the modification of a microfluidic, multi-chamber system so that electrical stimulation can be used to achieve increased levels of myelination while maintaining control of the cell culture microenvironment.
KeywordsMicrofluidic device Myelination Electrical stimulation Oligodendrocyte
This work was funded by US Department of Defense USAMRMC/TATRC/USAMRAA contracts W81XWH-08-2-0192, W81XWH-09-2-0186, W81XWH-10-BCRP-IDEA, and Maryland Stem Cell Research Fund.
- Demerens, C., Stankoff, B., Logak, M., Anglade, P., Allinquant, B., Couraud, F., et al. (1996). Induction of myelination in the central nervous system by electrical activity. Proceedings of the National Academy of Sciences of the United States of America, 93(18), 9887–9892.PubMedCrossRefGoogle Scholar
- Hur, E. M., Yang, I. H., Kim, D. H., Byun, J., Saijilafu, Xu, W. L., et al. (2011). Engineering neuronal growth cones to promote axon regeneration over inhibitory molecules. Proceedings of the National Academy of Sciences of the United States of America, 108(12), 5057–5062. doi:10.1073/pnas.1011258108.PubMedCrossRefGoogle Scholar
- Kimpinski, K., Campenot, R. B., & Mearow, K. (1997). Effects of the neurotrophins nerve growth factor, neurotrophin-3, and brain-derived neurotrophic factor (BDNF) on neurite growth from adult sensory neurons in compartmented cultures. Journal of Neurobiology, 33(4), 395–410. doi:10.1002/(SICI)1097-4695(199710)33:4<395:AID-NEU5>3.0.CO;2-5.PubMedCrossRefGoogle Scholar
- Li, Q., Brus-Ramer, M., Martin, J. H., & McDonald, J. W. (2010). Electrical stimulation of the medullary pyramid promotes proliferation and differentiation of oligodendrocyte progenitor cells in the corticospinal tract of the adult rat. Neuroscience Letters, 479(2), 128–133. doi:10.1016/j.neulet.2010.05.043.PubMedCrossRefGoogle Scholar
- Mehta, N. R., Lopez, P. H., Vyas, A. A., & Schnaar, R. L. (2007). Gangliosides and Nogo receptors independently mediate myelin-associated glycoprotein inhibition of neurite outgrowth in different nerve cells. Journal of Biological Chemistry, 282(38), 27875–27886. doi:10.1074/jbc.M704055200.PubMedCrossRefGoogle Scholar
- Vazdarjanova, A., Ramirez-Amaya, V., Insel, N., Plummer, T. K., Rosi, S., Chowdhury, S., et al. (2006). Spatial exploration induces ARC, a plasticity-related immediate-early gene, only in calcium/calmodulin-dependent protein kinase II-positive principal excitatory and inhibitory neurons of the rat forebrain. Journal of Comparative Neurology, 498(3), 317–329. doi:10.1002/cne.21003.PubMedCrossRefGoogle Scholar