Experimental study of mouse alveolar macrophages cultured in vitro under different conditions
10.3969/j.issn.1000-484X.2024.06.001
- VernacularTitle:不同条件下小鼠肺泡巨噬细胞体外培养的实验研究
- Author:
Hongfang YUN
1
;
Hui JIANG
;
Chunxiao NIU
;
Jiyan ZHANG
Author Information
1. 军事医学研究院军事认知与脑科学研究所,北京 100850
- Keywords:
Alveolar macrophages;
In vitro culture;
Bronchoalveolar lavage fluid;
Bone marrow;
Fetal liver
- From:
Chinese Journal of Immunology
2024;40(6):1121-1125
- CountryChina
- Language:Chinese
-
Abstract:
Obiective:To compare characteristics of alveolar macrophages(AMs)or AM-like cells in vitro cultured from different progenitors in mice of different ages in presence of GM-CSF,TGF-β and PPAR-γ agonist rosiglitazone(in brief,GTR),according to a previously reported optimized culture procedure.Methods: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,mononuclear cells were seeded into 12-well plates and cultured in presence of GTR.Generated cells were subjected to flow cytometry analysis to analyze expressions of AM markers F4/80,CD11b,CD170 and CD11c.Results:After culture,almost all BALF-derived cells were round-shaped under light microscope,about half bone marrow-derived cells appeared round-shaped,and about 85% fetal liver-derived cells showed round-shaped morphology.Frequency of BALF-AMs from 4-week-old mice was nearly 90%.Frequency and number of BALF-AMs increased with age of mice.Number of BALF-AMs from 12-week-old mice was nearly 1×106.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.Number of bone marrow-derived AM-like cells increased with age of mice,and about 0.8×106 cells/well bone marrow-derived AM-like cells were produced from 12-week-old mice.Frequency of fetal liver-derived AM-like cells was around 84% and their number could reach 4×106 cells/well.Conclusion:Purity and number of AMs or AM-like cells in vitro cultured from different progenitors in mice of different ages vary dramatically.Detailed condition should be chosen according to specific study requirements.
