title: Isolation of enriched glial populations from post-mortem human CNS material by immuno-laser capture microdissection.
abstract: Isolating individual populations of cells from post-mortem (PM) central nervous system (CNS) tissue for transcriptomic analysis will provide important insights into the pathogenesis of neurodegenerative diseases. To date, research on individual CNS cell populations has been hindered by the availability of suitable PM material, unreliable sample preparation and difficulties obtaining individual cell populations. In this paper we report how rapid immunohistochemistry combined with laser capture microdissection (immuno-LCM) enables the isolation of specific cell populations from PM CNS tissue, thereby enabling the RNA profile of these individual cell types to be investigated. Specifically, we detail methods for isolating enriched glial populations (astrocytes, oligodendrocytes and microglia) and confirm this cell enrichment by polymerase chain reaction (PCR). In addition, the study details the numbers of each glial population required to obtain 50ng RNA, a suitable amount of starting material required to carry out microarray analysis that potentially may identify alterations of cell-specific genes and pathways associated with a range of neurodegenerative disorders.
authors.affiliation: Sheffield Institute for Translational Neuroscience, The University of Sheffield, 385A Glossop Road, Sheffield, UK.
authors.forename: R
authors.initials: R
authors.lastname: Waller
authors.forename: M N
authors.initials: MN
authors.lastname: Woodroofe
authors.forename: S
authors.initials: S
authors.lastname: Francese
authors.forename: P R
authors.initials: PR
authors.lastname: Heath
authors.forename: S B
authors.initials: SB
authors.lastname: Wharton
authors.forename: P G
authors.initials: PG
authors.lastname: Ince
authors.forename: B
authors.initials: B
authors.lastname: Sharrack
authors.forename: J E
authors.initials: JE
authors.lastname: Simpson
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001253:Astrocytes / Q000166:cytology / Q000502:physiology; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002490:Central Nervous System / Q000166:cytology*; D005260:Female; D006801:Humans; D061346:Laser Capture Microdissection / Q000295:instrumentation / Q000379:methods*; D008297:Male; D017628:Microglia / Q000166:cytology / Q000502:physiology; D008875:Middle Aged; D019636:Neurodegenerative Diseases / Q000473:pathology / Q000503:physiopathology; D009457:Neuroglia / Q000166:cytology* / Q000502:physiology; D009836:Oligodendroglia / Q000166:cytology / Q000502:physiology; D014015:Tissue Banks / Q000592:standards