Abstract:
:Extraction of mouse spinal motor neurons from transgenic mouse embryos recapitulating some aspects of neurodegenerative diseases like amyotrophic lateral sclerosis has met with limited success. 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. 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(2) to maintain optimal viability. Functional astrocytes and microglia and small gamma motor neurons can be purified at the same time. This protocol will be a powerful and reliable method to obtain motor neurons and glia to better understand mechanisms underlying spinal cord diseases.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Beaudet MJ,Yang Q,Cadau S,Blais M,Bellenfant S,Gros-Louis F,Berthod Fdoi
10.1038/srep16763subject
Has Abstractpub_date
2015-11-18 00:00:00pages
16763issn
2045-2322pii
srep16763journal_volume
5pub_type
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