Function of microglia in organotypic slice cultures.

Abstract

We have examined the functional characteristics of microglia in an environment where the cytoarchitecture of the brain is preserved. Using organotypic slice culture under serum-free conditions, microglia initially demonstrated a rounded morphology but after 10 days in vitro (DIV), microglia in the slice were highly branched. Stimulation of the microglia at 4 DIV with phorbol ester significantly increased the number of cells stained with nitroblue tetrazolium, an indicator of superoxide anion production, compared to non-stimulated conditions. At 10 DIV, superoxide anion production was significantly less than that seen at 4 DIV and no increase in production was seen with phorbol ester stimulation. Phagocytosis of fluorescent latex beads and chemotaxis of microglia in response to zymosan activated serum was also reduced at 10 DIV compared to 4 DIV. These experiments indicate that microglia at 4 DIV in tissue slice culture have functional characteristics that resemble microglia in primary culture. However, prolonged culture of the slices results in a return of the microglia to a ramified and functionally down-regulated state, reminiscent of an "in vivo"-like environment. The organotypic slice culture, thus, provides a useful model system to I examine the interactions of microglia with neurons and other glia in the normal and injured brain.