Generation of Primary Astrocyte Cultures Devoid of Contaminating Microglia
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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.
