Abstract
To fully understand how external biomolecular environment influences axon growth, a method that can easily quantify the extent of axon growth as well as locally control their biomolecular environment is critically needed. Here, we describe a microfluidic culture platform capable of isolating CNS axons from neuronal somata for localized biomolecular manipulation as well as providing linearly guided axon growths for simple and easy quantification of the axon growth length. The axon isolation and guidance capability combined with the multi-compartment configuration make this platform ideal for investigating and screening drugs or other molecular factors that promote axon growth as well as regeneration.
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References
Qiu J, Cai D, Filbin MT (2000) Glial inhibition of nerve regeneration in the mature mammalian CNS. Glia 29:166–174
Fitch M, Silver J (2008) CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. Exp Neurol 209:294–301
Park KK et al (2008) Promoting axon regeneration in the adult CNS by modulation of the PTEN/mTOR pathway. Science 322:963–966
Campenot RB (1977) Local control of neurite development by nerve growth factor. PNAS 74:4516–4519
Hartikka J, Hefti F (1988) Development of septal cholinergic neurons in culture: plating density and glial cells modulate effects of NGF on survival, fiber growth, and expression of transmitter-specific enzymes. J Neurosci 8: 2967–2985
Brewer G, Torricelli J, Evege E, Price P (1993) Optimized survival of hippocampal neurons in B27‐supplemented neurobasal™, a new serum‐free medium combination. J Neurosci Res 35:567–576
Goldberg JL et al (2002) Retinal ganglion cells do not extend axons by default: promotion by neurotrophic signaling and electrical activity. Neuron 33:689–702
Goldberg JL, Klassen MP, Hua Y, Barres BA (2002) Amacrine-signaled loss of intrinsic axon growth ability by retinal ganglion cells. Science 296:1860–1864
Ito D et al (2010) Minimum neuron density for synchronized bursts in a rat cortical culture on multi-electrode arrays. Neuroscience 171: 50–61
Winzeler AM et al (2011) The lipid sulfatide is a novel myelin-associated inhibitor of CNS axon outgrowth. J Neurosci 31:6481–6492
Taylor AM et al (2005) A microfluidic culture platforms for CNS axonal injury. regeneration and transport. Nat Methods 2: 599–605
Park J, Koito H, Li J, Han A (2009) Microfluidic compartmentalized co-culture platform for CNS axon myelination research. Biomed Microdevices 11:1145–1153
Park J, Koito H, Li J, Han A (2012) Multi-compartment neuron-glia co-culture platform for localized CNS axon-glia interaction study. Lab Chip 12:3296–3304
Park J, Kim S, Park SI, Choe Y, Li J, Han A (2014) A microchip for quantitative analysis of CNS axon growth under localized biomolecular treatments. J Neurosci Methods 221:166–174
Koito H, Li J (2009) Preparation of rat brain aggregate cultures for neuron and glia development studies. J Vis Exp 31:1304
Acknowledgments
This work was supported by the National Institutes of Health/National Institute of Mental Health grant #1R21MH085267 and by the National Institutes of Health/National Institute of Neurological Disorders and Stroke grant #NS060017.
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Park, J., Kim, S., Li, J., Han, A. (2014). Axon Length Quantification Microfluidic Culture Platform for Growth and Regeneration Study. In: Murray, A. (eds) Axon Growth and Regeneration. Methods in Molecular Biology, vol 1162. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0777-9_7
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DOI: https://doi.org/10.1007/978-1-4939-0777-9_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0776-2
Online ISBN: 978-1-4939-0777-9
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