Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells

Dendritic cell-like cells (Mo-DCs) generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used as tools to treat cancer patients (DC-vaccines). Because Mo-DCs have multiple antigen presentation-related functions, in...

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Bibliographic Details
Published in:Journal of immunological methods 2004-04, Vol.287 (1), p.79-90
Main Authors: Tasaki, Akira, Yamanaka, Naoki, Kubo, Makoto, Matsumoto, Kotaro, Kuroki, Hideo, Nakamura, Katsuya, Nakahara, Chihiro, Onishi, Hideya, Kuga, Hirotaka, Baba, Eishi, Tanaka, Masao, Morisaki, Takashi, Katano, Mitsuo
Format: Article
Language:English
Subjects:
Antigen Presentation - immunology
Biological and medical sciences
Cell Culture Techniques - methods
Cell Line, Tumor
Cell Lineage
Cell Movement
Cell Survival
Coculture Techniques
Collagen
Collagen matrix
Dendritic Cells - physiology
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Gels
Humans
IL-12 production
Interferon-gamma - secretion
Interleukin-12 - secretion
Microscopy, Fluorescence
Microscopy, Phase-Contrast
Microscopy, Video
Migration
Mo-DC
Molecular immunology
Monocytes - cytology
Phagocytosis
T-Lymphocytes - immunology
Techniques
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Summary:Dendritic cell-like cells (Mo-DCs) generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used as tools to treat cancer patients (DC-vaccines). Because Mo-DCs have multiple antigen presentation-related functions, including phagocytosis, migration, cytokine production, and T cell stimulation, establishment of a method for simultaneously evaluating the various functions of Mo-DCs is important. We developed a new in vitro three-dimensional two-layer collagen matrix culture model that consists of a collagen gel containing Mo-DCs as the lower layer and a collagen gel containing necrotic GCTM-1 tumor cells and/or T cells as the upper layer. We used this system to observe simultaneously multiple functions of Mo-DCs by phase-contrast or fluorescence microscopy and to assess IL-12 secretion during more than 2 weeks of culture. We also observed interactions between Mo-DCs and necrotic GCTM-1 or T cells on an individual cell basis by time-lapse videomicroscopy. In addition, we collected Mo-DCs from the collagen gels by collagenase treatment and analyzed the expression of antigen presentation-related molecules such as HLA-DR, CD80, CD83, and CD86 on Mo-DCs. This model may be a useful tool for evaluation of the various functions of Mo-DCs used as DC vaccines and for studies of the complex behaviors of Mo-DCs in vivo.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2004.01.014