Journal of Capital Medical University ›› 2015, Vol. 36 ›› Issue (4): 618-621.doi: 10.3969/j.issn.1006-7795.2015.04.021

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A method for preparation of single cell suspension to detect the infiltration of inflammatory cells in mouse aorta by flow cytometry

Tian Cui1, Ren Hualiang2, Nie Hao2, Wang Xia1, Jiang Xue3, Li Fangda2, Wang Hongxia1   

  1. 1. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;
    2. Vascular Center Peking Union Medical College Hospital, Beijing 100073, China;
    3. Department of Cardiology, Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China
  • Received:2015-03-20 Online:2015-08-21 Published:2015-07-17
  • Supported by:

    This study was supported by Capital Medical University 2015 School Fund (2015JS14).

Abstract:

Objective To set up an effective method using flow cytometry technology to detect the infiltration of inflammatory cells in murine aorta by preparation of single cell suspensions. Methods Osmotic mini pumps with angiotensin Ⅱ(1 000 ng·kg-1·min-1) were implanted subcutaneously into mice for 3 d. The mice were sacrificed immediately after anesthesia, and the thoracic and abdominal aorta was cut off immediately after it was taken out. The vascular tissue of mice was digested with the digestive enzyme to prepare single cell suspension for flow cytometry analysis. Results 5×105 cells/mL could be isolated from each vascular tissue of mouse, and more intact cells and better viability were found in the single cell suspension; The results of flow cytometry showed angiotension Ⅱ treatment increased the number of neutrophisl, T lymphocyte cells and macrophages in the aorta. Conclusion This method is a simple, effective, stable and reliable one for the preparation of single-cell suspension of murine aorta, which will help to study the infiltration of inflammatory cells in murine aorta.

Key words: murine aortas, single cell suspensions, flow cytometry technology

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