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Isolation and Identification of Interstitial Macrophages from the Lungs Using Different Digestion Enzymes and Staining Strategies
- Source :
- Macrophages ISBN: 9781493978366
- Publication Year :
- 2018
- Publisher :
- Springer New York, 2018.
-
Abstract
- Interstitial macrophages (IMs) are present in multiple organs. Although there is limited knowledge of the unique functional role IM subtypes play, macrophages, in general, are known for their contribution in homeostatic tissue maintenance and inflammation such as clearing pathogens and debris and secreting inflammatory mediators and growth factors. IM subtypes have been identified in the heart, skin, and gut, and more recently we identified three distinct IMs in the lung. IMs express on their surface high levels of MerTK, CD64, and CD11b, with differences in CD11c, CD206, and MHC II expression, and referred to the three pulmonary IM subtypes as IM1 (CD11cloCD206+MHCIIlo), IM2 (CD11cloCD206+MHCIIhi), and IM3 (CD11chiCD206loMHCIIhi). In this chapter, we highlight how to extract IMs from the lung using three different digestion enzymes: elastase, collagenase D, and Liberase TM. Of these three commonly used enzymes, Liberase TM was the most effective at IM extraction, particularly IM3. Furthermore, alternative staining strategies to identify IMs were examined, which included CD64, MerTK, F4/80, and Tim4. Thus, future studies highlighting the functional role of IM subtypes will help further our understanding of how tissue homeostasis is maintained and inflammatory conditions are induced and resolved.
- Subjects :
- 0301 basic medicine
Cell Culture Techniques
Thermolysin
CD11c
Inflammation
Biology
Article
Monocytes
Mice
03 medical and health sciences
Macrophages, Alveolar
medicine
Animals
Collagenases
Lung
Tissue homeostasis
CD11b Antigen
Pancreatic Elastase
c-Mer Tyrosine Kinase
Receptors, IgG
Elastase
Membrane Proteins
MERTK
CD11c Antigen
Cell biology
030104 developmental biology
medicine.anatomical_structure
Gene Expression Regulation
Integrin alpha M
biology.protein
Collagenase
medicine.symptom
medicine.drug
Subjects
Details
- ISBN :
- 978-1-4939-7836-6
- ISBNs :
- 9781493978366
- Database :
- OpenAIRE
- Journal :
- Macrophages ISBN: 9781493978366
- Accession number :
- edsair.doi.dedup.....13a29d99c1e0460f06bc374a36f008c7