3D Cell Co-culture System on Hydrogel Micro-Patterned Surface Fabricated by Photolithography

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  • Yoshimoto Keitaro
    Graduate School of Pure and Applied Sciences, University of Tsukuba
  • Kojima Ryota
    Graduate School of Pure and Applied Sciences, University of Tsukuba
  • Takahashi Emiko
    Graduate School of Pure and Applied Sciences, University of Tsukuba
  • Ichino Masahiro
    Graduate School of Pure and Applied Sciences, University of Tsukuba
  • Miyoshi Hirotoshi
    Graduate School of Pure and Applied Sciences, University of Tsukuba
  • Nagasaki Yukio
    Graduate School of Pure and Applied Sciences, University of Tsukuba Master School of Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba Satellite Laboratory, International Center for Materials Nanoarchitectonics (WPI-MANA), National Institute of Materials Science (NIMS), University of Tsukuba Department of Life Sciences, Graduate School of Art and Sciences, The University of Tokyo

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We present herein the 3D cell co-culture systems, which is based on the cell spheroid formation on cell-adhesion micro-patterned glass surfaces fabricated by silane coupling treatment and photoirradiation technique. The typical method for constructing cell-adhesion micro-patterned glass surface is as follows; a mixture of methacryloyl-ended PEG telechelics and Irgacure 2959 as the photoinitiator was dropped on a silanized glass surface, which was pretreated with 3-(trimethoxysilyl)propyl methacrylate, and then the irradiation with UV light (254 nm) using a patterned mask with 100 μm aligned cavities separated by 100 μm intervals (edge-to-edge distance) was carried out. When hepatic cells (human hepatic cancer cell line and fetal mouse primary liver cell (FMLC)) were seeded onto the constructed surface, a spheroid array of these liver cells were fabricated and these hepatic functions were higher than the hepatic cells cultured on monolayer state. In this method, the co-culture of spheroids with monolayer-state cells is easily constructed and the functions of the FMLC spheroid were significantly upregulated by co-culture with nonparenhymal liver cells as feeder-cell.

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