High yield extraction of pure spinal motor neurons, astrocytes and microglia from single embryo and adult mouse spinal cord.

TitleHigh yield extraction of pure spinal motor neurons, astrocytes and microglia from single embryo and adult mouse spinal cord.
Publication TypeJournal Article
Year of Publication2015
AuteursBeaudet, M-J, Yang, Q, Cadau, S, Blais, M, Bellenfant, S, Gros-Louis, F, Berthod, F
JournalSci Rep
Volume5
Pagination16763
Date Published2015 Nov 18
ISSN2045-2322
KeywordsAnimals, Astrocytes, Biomarkers, Cell Separation, Cell Survival, Cells, Cultured, Embryo, Mammalian, Gene Expression, Mice, Mice, Transgenic, Microglia, Motor Neurons, Phenotype, Spinal Cord
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.

DOI10.1038/srep16763
Alternate JournalSci Rep
PubMed ID26577180
PubMed Central IDPMC4649473