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CLC number: R392.12

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Received: 2005-01-27

Revision Accepted: 2005-07-11

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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.12 P.1176~1181

http://doi.org/10.1631/jzus.2005.B1176


Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes


Author(s):  TANG Ling-ling, ZHANG Zhe, ZHENG Jie-sheng, SHENG Ji-fang, LIU Ke-zhou

Affiliation(s):  Department of Infectious Disease, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China

Corresponding email(s):   lltang72@126.com, shengjf@mail.hz.zj.cn

Key Words:  Dendritic cell, Monocyte, Phenotype, Flow cytometry


TANG Ling-ling, ZHANG Zhe, ZHENG Jie-sheng, SHENG Ji-fang, LIU Ke-zhou. Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes[J]. Journal of Zhejiang University Science B, 2005, 6(12): 1176~1181.

@article{title="Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes",
author="TANG Ling-ling, ZHANG Zhe, ZHENG Jie-sheng, SHENG Ji-fang, LIU Ke-zhou",
journal="Journal of Zhejiang University Science B",
volume="6",
number="12",
pages="1176~1181",
year="2005",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B1176"
}

%0 Journal Article
%T Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes
%A TANG Ling-ling
%A ZHANG Zhe
%A ZHENG Jie-sheng
%A SHENG Ji-fang
%A LIU Ke-zhou
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 12
%P 1176~1181
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B1176

TY - JOUR
T1 - Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes
A1 - TANG Ling-ling
A1 - ZHANG Zhe
A1 - ZHENG Jie-sheng
A1 - SHENG Ji-fang
A1 - LIU Ke-zhou
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 12
SP - 1176
EP - 1181
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B1176


Abstract: 
Objective: This study is aimed at developing a simple and easy way to generate dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) in vitro. Methods: PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods (collecting the attached cell and removing the suspension cell). DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM-CSF) and 20 ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFα) 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions (MLR) were used to characterize DC function after TNFα or lipopolysaccharide (LPS) stimulations for 24 h. Results: After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CD1a, CD80 and CD86, features of DCs. TNFα treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity. Conclusion: This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.

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