Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 114, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/54085
Keywords
Neuroscience; Issue 114; DRG isolation; pre-stretch; anisotropy; axon alignment; Schwann cell; myelination
Categories
Funding
- National Science Foundation [CBET 0941055, CBET 1510895]
- National Institute of Health [R21CA176854, R01GM089866, R01EB014986]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1510895] Funding Source: National Science Foundation
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Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to bridge the lesion site, properly organized axonal alignment is required. Since demyelination after nerve injury strongly impairs the conductive capacity of surviving axons, remyelination is critical for successful functioning of regenerated nerves. Previously, we demonstrated that mesenchymal stem cells (MSCs) aligned on a pre-stretch induced anisotropic surface because the cells can sense a larger effective stiffness in the stretched direction than in the perpendicular direction. We also showed that an anisotropic surface arising from a mechanical pre-stretched surface similarly affects alignment, as well as growth and myelination of axons. Here, we provide a detailed protocol for preparing a pre-stretched anisotropic surface, the isolation and culture of dorsal root ganglion (DRG) neurons on a pre-stretched surface, and show the myelination behavior of a co-culture of DRG neurons with Schwann cells (SCs) on a pre-stretched surface.
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