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However, astrocytes obtained from new born pups do not possess all the functional properties of adult astrocytes since they do not have processes and have a flat square-like shape instead of ramified stellar shape, and their gene expression profile and proliferating rate differs significantly from adult in vivo astrocytes that participates in versatile physiological processes in the central nervous system.
Consequently, astrocytes have to be screened for a large variety of markers to be adequately characterized in vitro.
Furthermore, MN cultures are particularly important in the study of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), which is characterized by MNs degeneration.
However, the mechanism of MN degeneration still remains unresolved despite the discovery that mutations in the superoxide-dismutase (SOD1) caused the disease in 1–2% of ALS patients, and the development of transgenic mice overexpressing the mutated protein (SOD1G93A and SOD1G37R mice) and recapitulating the disease symptoms.
In order to allow the culture of an adequate number of MNs expressing the desired genotype from mouse embryos, it would therefore be necessary to greatly increase MNs enrichment yields and to ultimately process each spinal cord collected from single embryo individually.
Attempts have been previously described for time-consuming MNs extraction protocols from single embryos using immune-affinity purification.
These protocols typically use a p75 (NTR)-antibody-based cell-sorting panning technique, p75 being an extracellular protein exclusively expressed in the spinal cord by MNs.
In these mouse models, massive death of motor neurons in the ventral horn of the spinal cord and loss of myelinated axons in ventral motor roots can be observed, ultimately leading to paralysis, muscle atrophy and death.
The easiest way to obtain MNs from mice is to collect them from embryos between the 12.