Document Detail


Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells.
MedLine Citation:
PMID:  19946957     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of Zhejiang University. Science. B     Volume:  10     ISSN:  1862-1783     ISO Abbreviation:  J Zhejiang Univ Sci B     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-30     Completed Date:  2010-03-01     Revised Date:  2010-09-27    
Medline Journal Info:
Nlm Unique ID:  101236535     Medline TA:  J Zhejiang Univ Sci B     Country:  China    
Other Details:
Languages:  eng     Pagination:  928-32     Citation Subset:  IM    
Affiliation:
Department of Renal Transplantation, Center of Nephropathy, the First Affiliated Hospital, Medicine College, Xi'an Jiaotong University, Xi'an 710061, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
CD4-Positive T-Lymphocytes / cytology*
Cell Separation
Cell Survival
Flow Cytometry / methods*
Immune System
Immunosuppressive Agents / pharmacology
Interleukin-2 Receptor alpha Subunit / biosynthesis*
Lymphocytes / cytology
Magnetics
Mice
Mice, Inbred BALB C
Chemical
Reg. No./Substance:
0/Immunosuppressive Agents; 0/Interleukin-2 Receptor alpha Subunit
Comments/Corrections

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