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. 2017 Jul 11:(125):55856.
doi: 10.3791/55856.

Spinal Cord Neurons Isolation and Culture from Neonatal Mice

Mohamed Eldeiry  1 , Katsuhiro Yamanaka  1 , T Brett Reece  1 , Muhammad Aftab  2
Affiliations

Spinal Cord Neurons Isolation and Culture from Neonatal Mice

Mohamed Eldeiry et al. J Vis Exp. .

Abstract

We present a protocol for the isolation and culture of spinal cord neurons. The neurons are obtained from neonatal C57BL/6 mice and are isolated on postnatal day 1-3. A mouse litter, usually 4-10 pups born from one breeding pair, is gathered for one experiment, and spinal cords are collected individually from each mouse after euthanasia with isoflurane. The spinal column is dissected out and then the spinal cord is released from the column. The spinal cords are then minced to increase the surface area of delivery for an enzymatic protease that allows for the neurons and other cells to be released from the tissue. Trituration is then used to release the cells into solution. This solution is subsequently fractionated in a density gradient to separate the various cells in solution, allowing for neurons to be isolated. Approximately 1-2.5 x 106 neurons can be isolated from one litter group. The neurons are then seeded onto wells coated with adhesive factors that allow for proper growth and maturation. The neurons take approximately 7 days to reach maturity in the growth and culture medium and can be used thereafter for treatment and analysis.

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References

    1. Qayumi AK, et al. Animal model for investigation of spinal cord injury caused by aortic cross-clamping. J Invest Surg. 1997;10(1-2):47–52. - PubMed
    1. Fang B, et al. Ischemic preconditioning protects against spinal cord ischemia-reperfusion injury in rabbits by attenuating blood spinal cord barrier disruption. Int J Mol Sci. 2013;14(5):10343–10354. - PMC - PubMed
    1. Taira Y, Marsala M. Effect of proximal arterial perfusion pressure on function, spinal cord blood flow, and histopathologic changes after increasing intervals of aortic occlusion in the rat. Stroke. 1996;27(10):1850–1858. - PubMed
    1. Stoppini L, Buchs P-A, Muller D. A simple method for organotypic cultures of nervous tissue. J Neurosci Methods. 1991;37(2):173–182. - PubMed
    1. Ahlemeyer B, Baumgart-Vogt E. Optimized protocols for the simultaneous preparation of primary neuronal cultures of the neocortex, hippocampus and cerebellum from individual newborn (P0. 5) C57Bl/6J mice. J Neurosci Methods. 2005;149(2):110–120. - PubMed

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