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Preparation of Single Cell Suspensions from Mouse Aorta
Desheng Hu1, Changjun Yin2, Sarajo K Mohanta2
1Institute for Immunology, Ludwig-Maximilians-University Munich, Munich, Germany; Institute of Molecular Immunology, Helmholtz Zentrum Munich, Munich, Germany.
Bio-Protocol
|June 24, 2016
Summary
This study presents a new protocol for isolating mouse aorta cells, crucial for understanding atherosclerosis. The method uses immunohistochemistry and fluorescence-activated cell sorting (FACS) to accurately quantify immune cells in aortic tissue.
Area of Science:
- Immunology
- Cardiovascular Biology
- Cell Biology
Background:
- Atherosclerosis is a chronic inflammatory arterial disease involving immune cell infiltration and plaque formation.
- Existing methods for analyzing aorta immune cells yield conflicting results due to variations in protocols and tissue contamination.
- Accurate quantification of immune cells within the aorta is essential for understanding disease mechanisms.
Purpose of the Study:
- To develop a robust protocol for isolating pure single-cell suspensions from mouse aortas.
- To enable accurate quantification and characterization of immune cells within the aortic wall.
- To address methodological challenges in previous studies, such as contamination from surrounding tissues.
Main Methods:
- A detailed protocol for isolating single cells from mouse aorta is described.
- The method emphasizes meticulous dissection to remove surrounding adipose tissue and lymph nodes.
- Combination of immunohistochemical staining and fluorescence-activated cell sorting (FACS) for cell quantification.
Main Results:
- The proposed protocol facilitates the isolation of bona fide aorta cells, minimizing contamination.
- Enables reliable quantification of diverse immune cell populations within the aorta.
- Provides a foundation for reproducible studies on immune cell roles in atherosclerosis.
Conclusions:
- This protocol offers a standardized approach for analyzing aorta-resident immune cells.
- Improved accuracy in immune cell quantification will advance our understanding of atherosclerosis.
- The method is critical for resolving discrepancies in current research and facilitating future discoveries.
