Isolation of enriched glial populations from post-mortem human CNS material by immuno-laser capture microdissection.

WALLER, R., WOODROOFE, M. Nicola, FRANCESE, Simona, HEATH, P.R., WHARTON, S.B., INCE, P.G., SHARRACK, B. and SIMPSON, J.E. (2012). Isolation of enriched glial populations from post-mortem human CNS material by immuno-laser capture microdissection. Journal of Neuroscience Methods, 208 (2), 108-113.

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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 50 ng 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.

Item Type:	Article
Research Institute, Centre or Group - Does NOT include content added after October 2018:	Biomedical Research Centre
Identification Number:	https://doi.org/10.1016/j.jneumeth.2012.04.014
Page Range:	108-113
Depositing User:	Rebecca Jones
Date Deposited:	15 May 2012 11:01
Last Modified:	18 Mar 2021 20:30
URI:	https://shura.shu.ac.uk/id/eprint/5058

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