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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2009, Vol. 10 Issue (12): 928-932    DOI: 10.1631/jzus.B0920205
Biomedicine     
Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells
Hang YAN, Chen-guang DING, Pu-xun TIAN, Guan-qun GE, Zhan-kui JIN, Li-ning JIA, Xiao-ming DING, Xiao-ming PAN, Wu-jun XUE
Department of Renal Transplantation, Center of Nephropathy, the First Affiliated Hospital, Medicine College, Xi\'an Jiaotong University, Xi\'an 710061, China
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Abstract  Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4+ CD25+ Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4+ CD25+ Treg cells in the in vitro proliferation experiments. Results: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4+ CD25+ Treg cells, however, obtained with both methods have similar immunosuppressive capacities. Conclusion: The result suggests that both methods can be used in isolating CD4+ CD25+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.

Key words： CD4+ CD25+ Treg cells      Flow cytometry sorting      Magnetic cell sorting     
Received: 14 July 2009     
CLC:  Q291  
  R392  
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Wu-jun XUE
Hang YAN
Chen-guang DING
Pu-xun TIAN
Guan-qun GE
Zhan-kui JIN
Li-ning JIA
Xiao-ming DING
Xiao-ming PAN
Cite this article:

Hang YAN, Chen-guang DING, Pu-xun TIAN, Guan-qun GE, Zhan-kui JIN, Li-ning JIA, Xiao-ming DING, Xiao-ming PAN, Wu-jun XUE. Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2009, 10(12): 928-932.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B0920205     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2009/V10/I12/928

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