title: High yield extraction of pure spinal motor neurons, astrocytes and microglia from single embryo and adult mouse 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.
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: Marie-Josée
authors.initials: MJ
authors.lastname: Beaudet
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: Qiurui
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Cadau
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Blais
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Bellenfant
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: François
authors.initials: F
authors.lastname: Gros-Louis
authors.affiliation: Centre LOEX de l'Université Laval, Centre de recherche du CHU de Québec, Québec (Québec), Canada.|Département de Chirurgie, Faculté de Médecine, Université Laval, Québec (Québec), Canada.
authors.forename: François
authors.initials: F
authors.lastname: Berthod
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D015415:Biomarkers; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D051379:Mice; D008822:Mice, Transgenic; D017628:Microglia / Q000166:cytology* / Q000378:metabolism; D009046:Motor Neurons / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D013116:Spinal Cord / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers