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
Hang YanWu-jun Xue

Abstract

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. 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. 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. 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.

References

Dec 5, 2006·Nature Medicine·Karin LoserStefan Beissert
Jan 5, 2007·Proceedings of the National Academy of Sciences of the United States of America·Joshua D MilnerWilliam E Paul
Mar 23, 2007·Nature·Masahiro OnoShimon Sakaguchi
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Jan 14, 2009·Translational Research : the Journal of Laboratory and Clinical Medicine·Guliang XiaXunrong Luo

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Citations

Sep 2, 2014·Wiley Interdisciplinary Reviews. Developmental Biology·Leo OtsukiAndrea H Brand
Dec 22, 2010·Biochemical and Biophysical Research Communications·Ping FanNan Hu
Jul 16, 2016·Immunological Investigations·Maryam AzimiEisa Salehi
Apr 5, 2019·Neurochemical Research·Klára JirákováPavla Jendelová
Dec 1, 2018·Nature Communications·Noelia DíazJuan M Vaquerizas
Nov 20, 2019·Bio-protocol·Loïc BinanSantiago Costantino

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