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


CONCLUSIONS
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.
Yan
Hang
H
Department of Renal Transplantation, Center of Nephropathy, the First Affiliated Hospital, Medicine College, Xi'an Jiaotong University, Xi'an 710061, China.
Ding
Chen-guang
CG
Tian
Pu-xun
PX
Ge
Guan-qun
GQ
Jin
Zhan-kui
ZK
Jia
Li-ning
LN
Ding
Xiao-ming
XM
Pan
Xiao-ming
XM
Xue
Wu-jun
WJ
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology*; D002469:Cell Separation; D002470:Cell Survival; D005434:Flow Cytometry / Q000379:methods*; D007107:Immune System; D007166:Immunosuppressive Agents / Q000494:pharmacology; D053645:Interleukin-2 Receptor alpha Subunit / Q000096:biosynthesis*; D008214:Lymphocytes / Q000166:cytology; D008280:Magnetics; D051379:Mice; D008807:Mice, Inbred BALB C
D007166:Immunosuppressive Agents; D053645:Interleukin-2 Receptor alpha Subunit