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Furthermore, extraction and long-term culture of adult mouse spinal motor neurons and glia remain also challenging.
We present here a protocol designed to extract and purify high yields of motor neurons and glia from individual spinal cords collected on embryos and adult (5-month-old) normal or transgenic mice.
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This protocol will be a powerful and reliable method to obtain motor neurons and glia to better understand mechanisms underlying spinal cord diseases.
This method is based on mild digestion of tissue followed by gradient density separation allowing to obtain two millions motor neurons over 92% pure from one E14.5 single embryo and more than 30,000 from an adult mouse.
These cells can be cultured more than 14 days in vitro at a density of 100,000 cells/cm to maintain optimal viability.
In vitro culture of purified motor neurons (MNs) is a useful approach to investigate numerous aspects of neuronal behavior, such as developmental biology, cell physiology, genomic, susceptibility to neurotoxins, etc.
Furthermore, MN cultures are particularly important in the study of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), which is characterized by MNs degeneration.