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Updated: Dec 31, 2025
Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages
Published on: April 20, 2018
Isolation and Long-term Cultivation of Mouse Alveolar Macrophages
Clara Jana-Lui Busch1,2, Jérémy Favret1,2,3, Laufey Geirsdóttir3
1Center for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Fetscherstraße 105, Dresden, Germany.
Abstract:
Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different conditions of BAL to obtain high yields of murine AM for in vitro culture and expansion of AM. In addition, we describe specific culture conditions, under which AM proliferate long-term in liquid culture in the presence of granulocyte-macrophage colony-stimulating factor. This method can be used to obtain large numbers of AM for in vivo transplantation or for in vitro experiments with primary mouse macrophages.
Insights
Researchers optimized bronchoalveolar lavage (BAL) and culture methods to yield large numbers of alveolar macrophages (AMs) for research. This protocol enables expansion of primary mouse AMs for in vitro and in vivo studies.
Area of Science:
- Immunology
- Cell Biology
- Pulmonology
Background:
- Alveolar macrophages (AMs) are crucial lung-resident immune cells that self-maintain without monocyte contribution.
- AMs reside in pulmonary alveoli and are typically harvested via bronchoalveolar lavage (BAL).
- Obtaining sufficient AMs for research can be challenging.
Purpose of the Study:
- To optimize bronchoalveolar lavage (BAL) techniques for high-yield murine alveolar macrophage (AM) collection.
- To establish robust in vitro culture conditions for long-term AM expansion.
- To provide a scalable method for generating primary mouse AMs for experimental use.
Main Methods:
- Comparison of various BAL conditions to maximize AM yield.
- Development of specific liquid culture protocols utilizing granulocyte-macrophage colony-stimulating factor (GM-CSF).
- Assessment of AM proliferation and viability under optimized culture conditions.
Main Results:
- Identified optimal BAL parameters for efficient murine AM harvesting.
- Demonstrated long-term proliferation of AMs in liquid culture with GM-CSF.
- Achieved high yields of viable AMs suitable for further experimentation.
Conclusions:
- Optimized BAL and culture methods provide a reliable source of murine AMs.
- This protocol facilitates in vitro and in vivo research using primary mouse macrophages.
- Enables large-scale generation of AMs for transplantation and experimental studies.
