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 Onishi, Hirotaka Kuga, Eishi Baba, Masao Tanaka, Takashi Morisaki, Mitsuo Katano

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16 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)79-90
Number of pages12
JournalJournal of Immunological Methods
Volume287
Issue number1-2
DOIs
Publication statusPublished - Apr 1 2004

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All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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