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

Akira Tasaki, Naoki Yamanaka, Makoto Kubo, Kotaro Matsumoto, Hideo Kuroki, Katsuya Nakamura, Chihiro Nakahara, Hideya Ohnishi, Hirotaka Kuga, Eishi Baba, Masao Tanaka, Takashi Morisaki, Mitsuo Katano

研究成果: ジャーナルへの寄稿記事

16 引用 (Scopus)

抄録

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.

元の言語英語
ページ(範囲)79-90
ページ数12
ジャーナルJournal of Immunological Methods
287
発行部数1-2
DOI
出版物ステータス出版済み - 4 1 2004

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Dendritic Cells
Monocytes
Collagen
Gels
Antigen Presentation
T-Lymphocytes
Vaccines
Phase-Contrast Microscopy
Video Microscopy
HLA-DR Antigens
Collagenases
Interleukin-12
Granulocyte-Macrophage Colony-Stimulating Factor
Fluorescence Microscopy
Phagocytosis
Interleukin-4
Neoplasms
Cytokines
Therapeutics

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

これを引用

Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells. / Tasaki, Akira; Yamanaka, Naoki; Kubo, Makoto; Matsumoto, Kotaro; Kuroki, Hideo; Nakamura, Katsuya; Nakahara, Chihiro; Ohnishi, Hideya; Kuga, Hirotaka; Baba, Eishi; Tanaka, Masao; Morisaki, Takashi; Katano, Mitsuo.

:: Journal of Immunological Methods, 巻 287, 番号 1-2, 01.04.2004, p. 79-90.

研究成果: ジャーナルへの寄稿記事

Tasaki, A, Yamanaka, N, Kubo, M, Matsumoto, K, Kuroki, H, Nakamura, K, Nakahara, C, Ohnishi, H, Kuga, H, Baba, E, Tanaka, M, Morisaki, T & Katano, M 2004, 'Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells', Journal of Immunological Methods, 巻. 287, 番号 1-2, pp. 79-90. https://doi.org/10.1016/j.jim.2004.01.014
Tasaki, Akira ; Yamanaka, Naoki ; Kubo, Makoto ; Matsumoto, Kotaro ; Kuroki, Hideo ; Nakamura, Katsuya ; Nakahara, Chihiro ; Ohnishi, Hideya ; Kuga, Hirotaka ; Baba, Eishi ; Tanaka, Masao ; Morisaki, Takashi ; Katano, Mitsuo. / Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells. :: Journal of Immunological Methods. 2004 ; 巻 287, 番号 1-2. pp. 79-90.
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abstract = "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.",
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AU - Kuroki, Hideo

AU - Nakamura, Katsuya

AU - Nakahara, Chihiro

AU - Ohnishi, Hideya

AU - Kuga, Hirotaka

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AU - Tanaka, Masao

AU - Morisaki, Takashi

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