title: Properties of human central nervous system neurons in a glia-depleted (isolated) culture system.
abstract: BACKGROUND
  Current methods for studying human neurons depend on a feeder layer of astroglia supplemented with animal serum to support the growing neurons. These requirements undermine many of the advantages provided by in vitro cell culture approaches when compared with more complex in vivo techniques.
  
  
  METHODS
  Here, we identified a reliable marker (MHCI) that allows for direct isolation of primary neurons from fetal human brain. We utilized a magnetic labeling and isolation technique to separate neurons from other neural cells. We established a defined condition, omitting the astroglial supports that could maintain the human neurons for varying amounts of time.
  
  
  RESULTS
  We showed that the new method significantly improved the purity of human neurons in culture without the need for further chemical/mechanical enrichment. We demonstrated the suitability of these neurons for functional studies including Rho-kinase dependent regulation of neurite outgrowth and ensheathment in co-cultures with oligodendrocyte progenitor cells derived from fetal human brain.
  
  
  METHODS
  The accountability for neuron-only seeding and the controllable density allows for better neuronal maturation and better visualization of the different neuronal compartments. The higher purity culture constitutes an effective system to study and screen for compounds that impact neuron biology without potential confounding effects from glial crowding.
  
  
  CONCLUSIONS
  High purity human neurons generated using the improved method will enable enhanced reliability in the discovery and development of drugs with neuroregenerative and neuroprotective activity.
authors.affiliation: Neuroimmunlogy Unit, Montreal Neurological Institute and Hospital, McGill University, Montreal, Quebec, H3A 2B4, Canada. Electronic address: sooyuen.leong@gmail.com.
authors.forename: Soo Yuen
authors.initials: SY
authors.lastname: Leong
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Kaplan
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec, H3G 1Y6, Canada.
authors.forename: Li-Chun
authors.initials: LC
authors.lastname: Wang
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec, H3G 1Y6, Canada.
authors.forename: Guillermina
authors.initials: G
authors.lastname: Almazan
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Alyson E
authors.initials: AE
authors.lastname: Fournier
authors.affiliation: Neuroimmunlogy Unit, Montreal Neurological Institute and Hospital, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Jack
authors.initials: J
authors.lastname: Antel
mesh_terms: D001921:Brain / Q000166:cytology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005333:Fetus; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism; D016501:Neurites / Q000502:physiology; D009457:Neuroglia / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D009836:Oligodendroglia / Q000502:physiology; D010799:Phycoerythrin / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013234:Stem Cells / Q000502:physiology; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D015395:Histocompatibility Antigens Class I; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D014404:Tubulin; D016875:tau Proteins; D010799:Phycoerythrin
keywords: Biomarker; Central nervous system; Human; Primary neurons