CLC number: Q291; R392
On-line Access: 2024-08-27
Received: 2023-10-17
Revision Accepted: 2024-05-08
Crosschecked: 2009-11-17
Cited: 10
Clicked: 6348
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[J]. Journal of Zhejiang University Science B, 2009, 10(12): 928-932.
@article{title="Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells",
author="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",
journal="Journal of Zhejiang University Science B",
volume="10",
number="12",
pages="928-932",
year="2009",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0920205"
}
%0 Journal Article
%T Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells
%A Hang YAN
%A Chen-guang DING
%A Pu-xun TIAN
%A Guan-qun GE
%A Zhan-kui JIN
%A Li-ning JIA
%A Xiao-ming DING
%A Xiao-ming PAN
%A Wu-jun XUE
%J Journal of Zhejiang University SCIENCE B
%V 10
%N 12
%P 928-932
%@ 1673-1581
%D 2009
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0920205
TY - JOUR
T1 - Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells
A1 - Hang YAN
A1 - Chen-guang DING
A1 - Pu-xun TIAN
A1 - Guan-qun GE
A1 - Zhan-kui JIN
A1 - Li-ning JIA
A1 - Xiao-ming DING
A1 - Xiao-ming PAN
A1 - Wu-jun XUE
J0 - Journal of Zhejiang University Science B
VL - 10
IS - 12
SP - 928
EP - 932
%@ 1673-1581
Y1 - 2009
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0920205
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.
[1] Campbell, D.J., Ziegler, S.F., 2007. FOXP3 modifies the phenotypic and functional properties of regulatory T cells. Nature Reviews Immunology, 7(4):305-310.
[2] Carrigan, S.O., Yang, Y.J., Issekutz, T., Forward, N., Hoskin, D., Johnston, B., Lin, T.J., 2009. Depletion of natural CD4(+)CD25(+) T regulatory cells with anti-CD25 antibody does not change the course of Pseudomonas aeruginosa-induced acute lung infection in mice. Immunobiology, 214(3):211-222.
[3] Golshayan, D., Jiang, S.P., Tsang, J., Garin, M.I., Mottet, C., Lechler, R.I., 2007. In vitro-expanded donor alloantigen-specific CD4(+)CD25(+) regulatory T cells promote experimental transplantation tolerance. Blood, 109(2): 827-835.
[4] Loser, K., Mehling, A., Loeser, S., Apelt, J., Kuhn, A., Grabbe, S., Schwarz, T., Penninger, J.M., Beissert, S., 2006. Epidermal RANKL controls regulatory T-cell numbers via activation of dendritic cells. Nature Medicine, 12(12): 1372-1379.
[5] Lu, Y.Q., Huang, W.D., Cai, X.J., Gu, L.H., Mou, H.Z., 2008. Hypertonic saline resuscitation reduces apoptosis of intestinal mucosa in a rat model of hemorrhagic shock. Journal of Zhejiang University-SCIENCE B, 9(11):879-884.
[6] Milner, J.D., Ward, J.M., Keane-Myers, A., Paul, W.E., 2007. Lymphopenic mice reconstituted with limited repertoire T cells develop severe, multiorgan, Th2-associated inflammatory disease. Proceedings of the National Academy of Sciences of the United States of America, 104(2): 576-581.
[7] Ono, M., Yaguchi, H., Ohkura, N., Kitabayashi, I., Nagamura, Y., Nomura, T., Miyachi, Y., Tsukada, T., Sakaguchi, S., 2007. Foxp3 controls regulatory T-cell function by interacting with AML1/Runx1. Nature, 446(7136):685-689.
[8] Steger, U., Kingsley, C.L., Karim, M., Wood, K.J., 2006. CD25(+)CD4(+) regulatory T cells develop in mice not only during spontaneous acceptance of liver allografts but also after acute allograft rejection. Transplantation, 82(9): 1202-1209.
[9] Wang, H.J., Zhao, L., Sun, Z.Y., Sun, L.G., Zhang, B.J., Zhao, Y., 2006. A potential side effect of cyclosporin A: inhibition of CD4(+)CD25(+) regulatory T cells in mice. Transplantation, 82(11):1484-1492.
[10] Xia, G.L., Shah, M., Luo, X.R., 2009. Prevention of allograft rejection by amplification of Foxp3+CD4+CD25+ regulatory T cells. Translational Research, 153(2):60-70.
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