In JoVE (1)
Other Publications (1)
Articles by Hannah Y. Collins in JoVE
Isolation and Culture of Rodent Microglia to Promote a Dynamic Ramified Morphology in Serum-free Medium Hannah Y. Collins1, Christopher J. Bohlen1 1Department of Neurobiology, Stanford University Efforts to understand microglial function in detail have been hindered by the lack of microglial culture models that recapitulate the properties of mature in vivo microglia. This protocol describes an isolation and culture approach designed to maintain robust survival of highly ramified mature rat microglia under defined-medium conditions.
Other articles by Hannah Y. Collins on PubMed
Diverse Requirements for Microglial Survival, Specification, and Function Revealed by Defined-Medium Cultures Neuron. May, 2017 | Pubmed ID: 28521131 Microglia, the resident macrophages of the CNS, engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol. Using microglial cultures that have never been exposed to serum, we demonstrate a dramatic and lasting change in phagocytic capacity after serum exposure. Finally, we find that mature microglia rapidly lose signature gene expression after isolation, and that this loss can be reversed by engrafting cells back into an intact CNS environment. These data indicate that the specialized gene expression profile of mature microglia requires continuous instructive signaling from the intact CNS.