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Updated: Jun 4, 2025
Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages
Published on: April 20, 2018
Protocol for isolating extracellular vesicles from alveolar macrophages phagocytosing MRSA in vitro cell culture
Songjie Bai1, Ruomeng Hu2, Xuehuan Wen2
1Department of Critical Care Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, China; Department of Cardiovascular Surgery, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, China.
Abstract:
Extracellular vesicles (EVs) play a crucial role in delivering bioactive cargo in infectious diseases. Here, we present a protocol for isolating EVs from alveolar macrophages (AMs) that phagocytose methicillin-resistant Staphylococcus aureus (MRSA) in vitro cell culture models. We describe steps for bacterial preparation; infection of AMs with MRSA; and isolation, purification, and characterization of EVs. This protocol provides a valuable perspective for studying EVs derived from pathogen-infected immune cells. For complete details on the use and execution of this protocol, please refer to Bai et al.1.
Insights
This study details a protocol for isolating extracellular vesicles (EVs) from macrophages infected with methicillin-resistant Staphylococcus aureus (MRSA). This method aids in understanding how EVs function in infectious diseases.
Area of Science:
- Microbiology
- Immunology
- Cell Biology
Background:
- Extracellular vesicles (EVs) are key mediators of intercellular communication, particularly in infectious diseases.
- Alveolar macrophages (AMs) are critical immune cells involved in lung defense against pathogens like Staphylococcus aureus.
- Understanding EV cargo in pathogen-infected immune cells is vital for developing novel therapeutic strategies.
Purpose of the Study:
- To establish a detailed protocol for the isolation and characterization of EVs from MRSA-infected AMs.
- To provide a reproducible method for researchers studying EV biogenesis and function in the context of bacterial infections.
- To facilitate the investigation of immune cell-derived EVs in infectious disease pathogenesis.
Main Methods:
- In vitro culture of alveolar macrophages (AMs).
- Infection of AMs with methicillin-resistant Staphylococcus aureus (MRSA).
- Isolation, purification, and characterization of extracellular vesicles (EVs) using established techniques.
Main Results:
- Successful isolation of EVs from MRSA-infected AMs.
- Characterization of the isolated EVs, confirming their identity and purity.
- Demonstration of a viable protocol for studying pathogen-host interactions via EVs.
Conclusions:
- The presented protocol enables the study of EVs derived from pathogen-infected immune cells.
- This methodology offers a valuable tool for investigating the role of EVs in infectious diseases.
- Further research can utilize this protocol to explore therapeutic targets related to EVs in MRSA infections.
