Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells

摘要

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 + CD25+ Treg cells%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of + CD25+ Treg cells%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>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 + CD25+ Treg cells%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>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 + CD25+ Treg cells%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>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 + CD25+ Treg cells%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>CD4+ CD25+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.