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Biomaterials. 2014 Mar;35(8):2428-35. doi: 10.1016/j.biomaterials.2013.12.014. Epub 2013 Dec 23.

Automatic fabrication of 3-dimensional tissues using cell sheet manipulator technique.

Author information

1
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan; CellSeed Incorporated, Katsura-Bldg. 4F, 3-61, Haramachi, Shinjuku-ku, Tokyo 162-0053, Japan.
2
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
3
ABLE Corporation, 7-9, Nishigoken-cho, Shinjyuku-ku, Tokyo 162-0812, Japan.
4
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Electronic address: tokano@abmes.twmu.ac.jp.

Abstract

Automated manufacturing is a key for tissue-engineered therapeutic products to become common-place and economical. Here, we developed an automatic cell sheet stacking apparatus to fabricate 3-dimensional tissue-engineered constructs exploiting our cell sheet manipulator technique, where cell sheets harvested from temperature-responsive culture dishes are stacked into a multilayered cell sheet. By optimizing the stacking conditions and cell seeding conditions, the apparatus was eventually capable of reproducibly making five-layer human skeletal muscle myoblast (HSMM) sheets with a thickness of approximately 70-80 μm within 100 min. Histological sections and confocal topographies of the five-layer HSMM sheets revealed a stratified structure with no delamination. In cell counts using trypsinization, the live cell numbers in one-, three- and five-layer HSMM sheets were equivalent to the seeded cell numbers at 1 h after the stacking processes; however, after subsequent 5-day static cultures, the live cell numbers of the five-layered HSMM sheets decreased slightly, while one- and three-layer HSMM sheets maintained their live cell numbers. This suggests that there are thickness limitations in maintaining tissues in a static culture. We concluded that by combining our cell sheet manipulator technique and industrial robot technology we can create a secure, cost-effective manufacturing system able to produce tissue-engineered products from cell sheets.

KEYWORDS:

3-D fabrication; Gelatin; Myoblast; Sheet; Thermally responsive material

[Indexed for MEDLINE]

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