Title

Phenotypic and Functional Characteristics of Dendritic Cells Derived from Human Peripheral Blood Monocytes

Authors

湯靈玲(Ling-Ling Tang);張哲(Zhe Zhang);鄭傑勝(Jie-Sheng Zheng);盛吉芳(Ji-Fang Sheng);劉克洲(Ke-Zhou Liu)

Key Words

Dendritic cell ; Monocyte ; Phenotype ; Flow cytometry

PublicationName

Journal of Zhejiang University Science

Volume or Term/Year and Month of Publication

6B卷12期(2005 / 12 / 15)

Page #

1176 - 1181

Content Language

英文

English 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 30ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM-CSF) and 20ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFα) 30ng/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.

Topic Category 基礎與應用科學 > 基礎與應用科學綜合
醫藥衛生 > 醫藥衛生綜合
生物農學 > 生物農學綜合
工程學 > 工程學綜合