Bone marrow-derived macrophage obtained from fresh or cryopreserved bone marrow cells respond similarly to the infection with <i>Leishmania amazonensis</i>.

<p>Bone marrow-derived macrophages (BMDM) obtained from fresh or cryopreserved (frozen) bone marrow cells were infected with <i>L. amazonensis</i> promastigotes constitutively expressing the GFP at an MOI of 10 parasites per cell. After 48 h and 96 h of infection, the percentages of <i>L. amazonensis</i>-GFP⁺ and propidium iodide-positive cells were estimated by flow cytometry. (A) Percentage of GFP⁺ BMDM obtained from fresh (white bars) and frozen (black bars) BM cells. (B) Mean fluorescence intensities (MIF) for GFP signal. (C) Representative images of cultures infected for 48 h e 96 h with <i>L. amazonensis</i> GFP. (D) FACS profiles of BMDM obtained from fresh and frozen BM cells infected with <i>L. amazonensis</i> GFP. Shown are the percentages of cells with signal for GFP and PI. (E) Bar graphics indicating the percentage of propidium iodide-positive cells in uninfected cultures (control), cells treated with 0.05% Tween 20 (Tween) or after infection with <i>L. amazonensis</i> for 48 and 96 hs. Data are representative of those found in three independent experiments. No statistically significant differences were detected between BMDM obtained from fresh or frozen BM cells (<i>P</i>>0.05).