Macrophage Populations Isolated From Sham and Saline Intratracheally Instilled Mice: Maturity and Phenotype Data

Published: 2 March 2021| Version 1 | DOI: 10.17632/n466k4xbws.1
Contributor:
Thea Golden

Description

Resident and recruited macrophages were identified via a chimeric mouse model. The macrophage populations of these chimeras were also assessed following sham and saline intratracheal instillation. Alveolar macrophages were collected via bronchoalveolar lavage and protein expression measured by flow cytometry. Macrophages were assessed for maturity and phenotype.

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C57/BL6 mice underwent irradiation followed by adoptive transfer of bone marrow cells. C57BL/6-Tg (CAG-EGFP)10sb/J bone marrow was used to identify bone marrow derived cells based on GFP expression. NOS2-/- mice were used to create bone marrow knockout and pulmonary knockout chimeras. Following 6 weeks of recovery, mice either underwent sham or saline intratracheal instillation. 8 and 15 days post instillation, mice underwent bronchoalveolar lavage and cells were counted via Mutisizer 3 cell counter. Cells were stained via manufacturer protocol with fluorescent conjugated antibodies; F4/80, Cd11b, Cd11c, Cd206 and Ly6C. Beckman Coulter Gallios Flow Cytometer was used to measure samples, and Kaluza software for analysis.