Fig. 7.

Enzymatic isolation of brain cells with papain and dispase II to evaluate glia response to LPS. Animals were injected with LPS (0.5 mg/Kg, i.p.) to induce a mild inflammatory response. After 3 days, mice were sacrificed. Brain cells were isolated using papain and dispase II combined with gentle mechanical dissociation and DNase II incubation. Purification with 30 % Percoll was followed by flow cytometry analysis of (A) resting microglia (CD11b+/CD45^low) and activated microglia/macrophages (CD11b+/CD45^high), or (B) astrocytes (ACSA-2+/CD45-). Values are expressed as percentage of total CD45+ cells (A) or CD45-cells (B) and correspond to mean ± S.E.M. (n = 7–9). *, p < 0.05. (C) Another set of control and LPS-treated animals was fixed by perfusion, and brain sections stained for immunofluorescence detection of either Iba-1 or GFAP as markers of microglia and astrocytes, respectively. Representative images containing the dentate gyrus of hippocampus are shown (scale bar, 100 μm).