A three-dimensional in vitro model of the peripheral nervous system
Source abstract
Abstract
Abstract Recent advances in three-dimensional (3D) cell culture models developed on organ-on-a-chip or microfluidic devices have shown their capability to recapitulate the in vivo microenvironment as well as their potential as tools in biomedical research. Here, we present an in vitro model of the peripheral nervous system (PNS) by establishing a coculture model of motor neurons (MNs) and Schwann cells (SCs) in a 3D environment in a microengineered extracellular matrix hydrogel scaffold. The collagen scaffold placed at the center of the microdevice provided a 3D cellular microenvironment where the axons of MNs were allowed to actively interact with SCs during their growth and maturation. By treating the MN–SC coculture model with ascorbic acid, we were able to model the myelination process in the PNS, which was evidenced by the increased expression of myelin markers in SCs. Moreover, we show that this can be reversed by treating myelinated nerve fibers with glial growth factor (neuregulin-1 isoform) to potentially block the formation of the myelin sheath and induce demyelination. Our 3D cell culture model may be used to achieve active control of the myelinating and demyelinating processes in the PNS and thus may offer new opportunities to study pathophysiological processes involved in motor neuron diseases by in vitro modeling.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Nerve injury and regeneration · 3D Printing in Biomedical Research · Cellular Mechanics and Interactions
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
References (45)
Show source-provided references
1. Garbay, B., Heape, A. M., Sargueil, F. & Cassagne, C. Myelin synthesis in the peripheral nervous system. Prog. Neurobiol. 61, 267–304 (2000).10.1016/S0301-0082(99)00049-0 ↗
2. Salzer, J. L. Schwann cell myelination. Cold Spring Harb. Perspect. Biol. 7, 1–26 (2015).10.1101/cshperspect.a020529 ↗
3. Jessen, K. R. & Mirsky, R. The origin and development of glial cells in peripheral nerves. Nat. Rev. Neurosci. 6, 671–682 (2005).10.1038/nrn1746 ↗
4. Salzer, J. L., Brophy, P. J. & Peles, E. Molecular domains of myelinated axons in the peripheral nervous system. Glia 56, 1532–1540 (2008).10.1002/glia.20750 ↗
5. Callizot, N., Combes, M., Steinschneider, R. & Poindron, P. A new long term in vitro model of myelination. Exp. Cell Res. 317, 2374–2383 (2011).10.1016/j.yexcr.2011.07.002 ↗
6. Stettner, M. et al. A reliable in vitro model for studying peripheral nerve myelination in mouse. J. Neurosci. Methods 214, 69–79 (2013).10.1016/j.jneumeth.2013.01.009 ↗
7. Hyung, S. et al. Coculture of primary motor neurons and Schwann cells as a model for in vitro myelination. Sci. Rep. 5, 15122 (2015).10.1038/srep15122 ↗
8. Shin, Y. et al. Microfluidic assay for simultaneous culture of multiple cell types on surfaces or within hydrogels. Nat. Protoc. 7, 1247–1259 (2012).10.1038/nprot.2012.051 ↗
9. van Duinen, V., Trietsch, S. J., Joore, J., Vulto, P. & Hankemeier, T. Microfluidic 3D cell culture: from tools to tissue models. Curr. Opin. Biotechnol. 35, 118–126 (2015).10.1016/j.copbio.2015.05.002 ↗
10. Eldridge, C. F., Bunge, M. B., Bunge, R. P. & Wood, P. M. Differentiation of axon-related Schwann cells in vitro. I. Ascorbic acid regulates basal lamina assembly and myelin formation. J. Cell Biol. 105, 1023–1034 (1987).10.1083/jcb.105.2.1023 ↗
11. Laszkiewicz, I., Wiggins, R. C. & Konat, G. Ascorbic acid upregulates myelin gene expression in C6 glioma cells. Metab. Brain Dis. 7, 157–164 (1992).10.1007/BF01000161 ↗
12. Zanazzi, G. et al. Glial growth factor/neuregulin inhibits Schwann cell myelination and induces demyelination. J. Cell Biol. 152, 1289–1299 (2001).10.1083/jcb.152.6.1289 ↗
13. Wiese, S. et al. Isolation and enrichment of embryonic mouse motoneurons from the lumar spinal cord of individual mouse embryos. Nat. Protoc. 5, 31–38 (2010).10.1038/nprot.2009.193 ↗
14. Toy, D. & Namgung, U. Role of glial cells in axonal regeneration. Exp. Neurobiol. 22, 68–76 (2013).10.5607/en.2013.22.2.68 ↗
15. Mitchel, J. A. & Hoffman-Kim, D. Cellular scale anisotropic topography guides Schwann cell motility. PLoS ONE 6, e24316 (2011).10.1371/journal.pone.0024316 ↗
16. Lopez-Fagundo, C., Bar-Kochba, E., Livi, L. L., Hoffman-Kim, D. & Frank, C. Three-dimensional traction forces of Schwann cells on compliant substrates. J. R. Soc. Interface 11, 20140247 (2014).10.1098/rsif.2014.0247 ↗
17. Edmondson, R., Broglie, J. J., Adcock, A. F. & Yang, L. Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay. Drug Dev. Technol. 12, 207–218 (2014).10.1089/adt.2014.573 ↗
18. Bunge, R. P. Tissue culture observations relevant to the study of axon–Schwann cell interactions during peripheral nerve development and repair. J. Exp. Biol. 132, 21–34 (1987).10.1242/jeb.132.1.21 ↗
19. Fawcett, J. W. & Keynes, R. J. Peripheral nerve regeneration. Annu. Rev. Neurosci. 13, 43–60 (1990).10.1146/annurev.ne.13.030190.000355 ↗
20. Bunge, R. P. The role of the Schwann cell in trophic support and regeneration. J. Neurol. 242, S19–S21 (1994).10.1007/BF00939235 ↗
21. Yoshimura, K. & Takeda, S. Hedgehog signaling regulates myelination in the peripheral nervous system through primary cilia. Differentiation 83, S78–S85 (2012).10.1016/j.diff.2011.10.006 ↗
22. Feltri, M. L., Poitelon, Y. & Previtali, S. C. How Schwann cells sort axons: new concepts. Neuroscientist 22, 252–265 (2016).10.1177/1073858415572361 ↗
23. Thompson, D. M. & Buettner, H. M. Neurite outgrowth is directed by Schwann cell alignment in the absence of other guidance cues. Ann. Biomed. Eng. 34, 669–676 (2006).10.1007/s10439-005-9053-9 ↗
24. Heermann, S. & Schwab, M. H. Molecular control of Schwann cell migration along peripheral axons. Cell Adhes. Migr. 7, 18–22 (2013).10.4161/cam.22123 ↗
25. Liazoghli, D., Roth, A. D., Thostrup, P. & Colman, D. R. Substrate micropatterning as a new in vitro cell culture system to study myelination. ACS Chem. Neurosci. 3, 90–95 (2012).10.1021/cn2000734 ↗
26. Rumsey, J. W. et al. Node of Ranvier formation on motoneurons in vitro. Biomaterials 30, 3567–3572 (2009).10.1016/j.biomaterials.2009.03.023 ↗
27. Poliak, S. & Peles, E. The local differentiation of myelinated axons at nodes of Ranvier. Nat. Rev. Neurosci. 4, 968–980 (2003).10.1038/nrn1253 ↗
28. Son, Y. J. & Thompson, W. J. Schwann cell processes guide regeneration of peripheral axons. Neuron 14, 125–132 (1995).10.1016/0896-6273(95)90246-5 ↗
29. Scheib, J. & Hoke, Ahmet Advances in peripheral nerve regeneration. Nat. Rev. Neurol. 9, 668–676 (2013).10.1038/nrneurol.2013.227 ↗
30. Martini, R. & Schachner, M. Molecular bases of myelin formation as revealed by investigations on mice deficient in glial cell surface molecules. Glia 19, 298–310 (1997).10.1002/(SICI)1098-1136(199704)19:4<298::AID-GLIA3>3.0.CO;2-U ↗
31. Eichberg, J. Myelin P0: new knowledge and new roles. Neurochem. Res. 27, 1331–1340 (2002).10.1023/A:1021619631869 ↗
32. Fex Svenningsen, A., Shan, W. S., Colman, D. R. & Pedraza, L. Rapid method for culturing embryonic neuron-glial cell cocultures. J. Neurosci. Res. 72, 565–573 (2003).10.1002/jnr.10610 ↗
33. Chan, J. R. et al. NGF controls axonal receptivity to myelination by Schwann cells or oligodendrocytes. Neuron 43, 183–191 (2004).10.1016/j.neuron.2004.06.024 ↗
34. Eshed, Y. et al. Gliomedin mediates Schwann cell-axon interaction and the molecular assembly of the nodes of Ranvier. Neuron 47, 215–229 (2005).10.1016/j.neuron.2005.06.026 ↗
35. Liu, N., Varma, S., Shooter, E. M. & Tolwani, R. J. Enhancement of Schwann cell myelin formation by K252a in the Trembler-J mouse dorsal root ganglion explant culture. J. Neurosci. Res. 79, 310–317 (2005).10.1002/jnr.20357 ↗
36. Honkanen, H. et al. Isolation, purification and expansion of myelination-competent, neonatal mouse Schwann cells. Eur. J. Neurosci. 26, 953–964 (2007).10.1111/j.1460-9568.2007.05726.x ↗
37. Gingras, M., Beaulieu, M. M., Gagnon, V., Durham, H. D. & Berthod, F. In vitro study of axonal migration and myelination of motor neurons in a three-dimensional tissue-engineered model. Glia 56, 354–364 (2008).10.1002/glia.20617 ↗
38. Paivalainen, S. et al. Myelination in mouse dorsal root ganglion/Schwann cell cocultures. Mol. Cell Neurosci. 37, 568–578 (2008).10.1016/j.mcn.2007.12.005 ↗
39. Rangaraju, S., Madorsky, I., Pileggi, J. G., Kamal, A. & Notterpek, L. Pharmacological induction of the heat shock response improves myelination in a neuropathic model. Neurobiol. Dis. 32, 105–115 (2008).10.1016/j.nbd.2008.06.015 ↗
40. Feinberg, K. et al. A glial signal consisting of gliomedin and NrCAM clusters axonal Na+ channels during the formation of nodes of Ranvier. Neuron 65, 490–502 (2010).10.1016/j.neuron.2010.02.004 ↗
41. Susuki, K. et al. Schwann cell spectrins modulate peripheral nerve myelination. Proc. Natl Acad. Sci. USA 108, 8009–8014 (2011).10.1073/pnas.1019600108 ↗
42. Rumsey, J. W. et al. Myelination and node of Ranvier formation on sensory neurons in a defined in vitro system. In Vitro Cell Dev. Biol. Anim. 49, 608–618 (2013).10.1007/s11626-013-9647-8 ↗
43. Huval, R. M. et al. Microengineered peripheral nerve-on-a-chip for preclinical physiological testing. Lab Chip 15, 2221–2232 (2015).10.1039/C4LC01513D ↗
44. Sakai, K., Shimba, K., Kotani, K. & Jimbo, Y. A co-culture microtunnel technique demonstrating a significant contribution of unmyelinated Schwann cells to the acceleration of axonal conduction in Schwann cell-regulated peripheral nerve development. Integr. Biol. 9, 678–686 (2017).10.1039/C7IB00051K ↗
45. Khoshakhlagh, P., Sivakumar, A., Pace, L. A., Sazer, D. W. & Moore, M. J. Methods for fabrication and evaluation of a 3D microengineered model of myelinated peripheral nerve. J. Neural. Eng. 15, 064001 (2018).10.1088/1741-2552/aae129 ↗