Experimental study of mouse alveolar macrophages cultured in vitro under different conditions

After bronchoalveolar lavage fluid (BALF) and bone marrow were isolated from mice of different ages and fetal livers were isolated from embryonic-day-15-17 mice, the mononuclear cells were seeded into 12-well plates and cultured in the presence of GTR. The generated cells were subjected to flow cytometry analysis to analyze the expression of AM markers.Results After culture, almost all BALF-derived cells were round-shaped under the light microscope, about half bone marrow-derived cells appeared round-shaped, and about 85% fetal liver-derived cells showed round-shaped morphology.

The frequency of BALF-AMs from 4-week-old mice was nearly 90%. The frequency and number of BALF-AMs increased with the age of mice. The number of BALF-AMs from 12-week-old mice was nearly 1 × 106.

The frequency of bone marrow-derived AM-like cells from 4-week-old mice was lower than 20% and increased to about 40-45% for 8-week-old and 12-week-old mice. The number of bone marrow-derived AM-like cells increased with the age of mice and about 0.8 × 106 bone marrow-derived AM-like cells/well were produced from 12-week-old mice. The frequency of fetal liver-derived AM-like cells was around 84% and their number could reach 4 × 106 /well.Conclusions The purity and number of AMs or AM-like cells in vitro cultured from different progenitors in mice of different ages vary dramatically.

The detailed condition should be chosen according to specific study requirements.