Department of Renal Transplantation, Center of Nephropathy, the First Affiliated Hospital, Medicine College, Xi'an Jiaotong University, Xi'an 710061, China.
J Zhejiang Univ Sci B. 2009 Dec;10(12):928-32. doi: 10.1631/jzus.B0920205.
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.
本文比较了两种细胞分选方法(磁珠分选和流式细胞术分选)在分离和功能分析小鼠 CD4+CD25+调节性 T(Treg)细胞中的应用,以进一步了解 Treg 细胞的功能。
我们分别采用磁珠分选和流式细胞术分选鉴定 CD4+CD25+Treg 细胞。磁珠分选后,我们进一步采用流式细胞术分析 CD4+CD25+Treg 细胞的纯度,台盼蓝染色检测细胞活力,碘化丙啶(PI)染色评估细胞活力。在体外增殖实验中检测 CD4+CD25+Treg 细胞的免疫抑制作用。
结果表明,与流式细胞术分选相比,磁珠分选需要更多的时间和精力,但获得的活细胞比流式细胞术分选少。然而,两种方法获得的 CD4+CD25+Treg 细胞具有相似的免疫抑制能力。
结果提示两种方法均可用于分离 CD4+CD25+Treg 细胞,可根据具体需求和方法的可获得性选择最佳方法。