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An in vitro Model of Neuron-macrophage Interaction to Generate Macrophages with Neurite Outgrowth Properties   

Edited by
Soyun Kim
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Original research article

A brief version of this protocol appeared in:
The Journal of Neuroscience
Dec 2015

Abstract

Macrophages are known to play beneficial roles in axon regeneration after nerve injury. To develop an in vitro model in which injury signals can elicit pro-regenerative macrophage activation, we established co-cultures consisting of adult dorsal root ganglia sensory neurons and peritoneal macrophages and added cAMP analogue dibutyryl cAMP. The conditioned medium collected from the co-cultures exhibited robust neurite outgrowth activities. The neurite outgrowth activities were almost completely abrogated by addition of minocycline, a macrophage deactivator, indicating that factors responsible for neurite outgrowth are produced by activated macrophages.

Background

CNS neurons of adult mammals do not spontaneously regenerate axons after injury. Preconditioning peripheral nerve injury allows the dorsal root ganglia (DRG) sensory axons to regenerate central branches by promoting expression of regeneration-associated genes. We have previously showed that activated macrophages in the DRG following preconditioning injury critically contribute to enhancement intrinsic regeneration capacity of the DRG sensory neuron (Kwon et al., 2013). To identify molecular factors involved in the activation of macrophages following nerve injury, we have developed an in vitro model in which neuron-macrophages interactions are elicited by cAMP, a well-known reagent to enhance regenerative capacity of neurons. Compared to the previous model to activate macrophages using zymosan, our model utilizes a more physiologic stimulus resembling molecular events in the preconditioning peripheral injury model.

Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Yun, H. J. and Kim, B. S. (2016). An in vitro Model of Neuron-macrophage Interaction to Generate Macrophages with Neurite Outgrowth Properties. Bio-protocol 6(22): e2012. DOI: 10.21769/BioProtoc.2012.
  2. Kwon, M. J., Shin, H. Y., Cui, Y., Kim, H., Thi, A. H., Choi, J. Y., Kim, E. Y., Hwang, D. H. and Kim, B. G. (2015). CCL2 mediumtes neuron-macrophage interactions to drive proregenerative macrophage activation following preconditioning injury. J Neurosci 35(48): 15934-15947.
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