Generation of primary astrocyte cultures devoid of contaminating microglia

Tracy F. Uliasz, Mary E. Hamby, Nicole A. Jackman, James A. Hewett, Sandra J. Hewett

Research output: Chapter in Book/Report/Conference proceedingChapter

16 Scopus citations

Abstract

Microglia, resident phagocytic cells of the central nervous system, are frequent contaminants of astrocyte cultures. Unfortunately and not always fully appreciated, contamination by microglia can confound results of studies designed to elucidate the molecular mechanisms underlying astrocyte-specific responses. The paradigm described herein employs the mitotic inhibitor, cytosine β-d-arabinofuranoside, followed by the lysosomotropic agent, leucine methylester, to maximally deplete microglia, thereby generating highly enriched astrocyte monolayers that remain viable and functional. Successful removal of microglia from confluent monolayers of primary astrocyte cultures is achieved without the need for cell passage and successful reduction is confirmed by depletion of microglial-specific markers.

Original languageEnglish (US)
Title of host publicationAstrocytes
Subtitle of host publicationMethods and Protocols
EditorsRichard Milner
Pages61-79
Number of pages19
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume814
ISSN (Print)1064-3745

Keywords

  • Astrocytes
  • Cell culture
  • Lysosomotropism
  • Microglia
  • Mitotic inhibition

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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