title: Isolation of living neurons from human elderly brains using the immunomagnetic sorting DNA-linker system.
abstract: Isolation and culture of mature neurons from affected brain regions during diseased states provide a well-suited in vitro model system to study age-related neurodegeneration under dynamic conditions at cellular levels. We have developed a novel technique to isolate living neurons from rapidly autopsied human elderly brains, and have succeeded in keeping them alive in vitro. Specifically, the parietal cortex blocks were fractionated by density gradients and further enriched for neurons by an immunomagnetic sorting DNA-linker technique. The postmortem interval averaged 2.6 hours. After isolation and purification of neurons using this technology, the cells were maintained in vitro for 2 weeks. Our evaluation revealed that 80% of the isolated cells were neurons and they exhibited neurotransmitter phenotypes (glutamate and gamma-aminobutyric acid) as well as glutamate receptors. Studies on cell viability and calcium influx suggest that these isolated living cortical neurons still retain their typical neuronal functions. Our present study demonstrates that neurons isolated from human elderly brain autopsies can survive in vitro and maintain their functional properties. Our study has opened an opportunity to apply such neurons to dynamic pharmacological studies of neurological disorders at the single-cell level.
authors.affiliation: Haldeman Laboratory of Molecular and Cellular Neurobiology, Sun Health Research Institute, Sun City, AZ 85351, USA.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Konishi
authors.forename: Kristina
authors.initials: K
authors.lastname: Lindholm
authors.forename: Li-Bang
authors.initials: LB
authors.lastname: Yang
authors.forename: Rena
authors.initials: R
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Shen
mesh_terms: D000368:Aged; D001921:Brain / Q000166:cytology*; D002118:Calcium / Q000032:analysis; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D009474:Neurons / Q000737:chemistry / Q000166:cytology*; D018377:Neurotransmitter Agents / Q000032:analysis; D010641:Phenotype
chemical_list: D018377:Neurotransmitter Agents; D004247:DNA; D002118:Calcium