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

机译：使用芯板机械手技术自动制造三维组织

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摘要

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 70-80μm within 100min. 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 1h 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.
机译：自动化制造是组织工程治疗产品成为共同和经济的关键。这里，我们开发了一种自动细胞片堆叠装置，用于制造利用我们的细胞片机械手技术的三维组织工程构建体，其中从温度响应培养皿收获的细胞片堆叠成多层细胞片。通过优化堆叠条件和细胞播种条件，该装置最终能够在100min范围内再现厚度为约70-80μm的五层人骨骼肌肌细胞（HSMM）片。五层HSMM纸张的组织学区和共聚焦拓扑表现出一种没有分层的分层结构。在使用胰蛋白酶化的细胞计数中，在堆叠过程之后的1小时内，单个，三层和五层HSMM片材中的活细胞数量相当于接种细胞数;然而，在随后的5天静态培养物之后，五层HSMM片材的活细胞数略微下降，而单层HSMM纸张保持其活细胞数。这表明在静态培养中保持组织存在厚度限制。我们得出结论，通过组合我们的蜂窝手柄技术和工业机器人技术，我们可以创建一种能够从细胞片生产组织工程产品的安全性成本效益的制造系统。

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  • 来源
    《Biomaterials》 |2014年第8期|共8页
  • 作者

    KikuchiT.; ShimizuT.; WadaM.; YamatoM.; OkanoT.;

  • 作者单位

    Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University (TWIns);

    Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University (TWIns);

    ABLE Corporation 7-9 Nishigoken-cho Shinjyuku-ku Tokyo 162-0812 Japan;

    Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University (TWIns);

    Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University (TWIns);

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物医学工程;
  • 关键词

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

    机译：3-D制作;明胶;肌细胞;纸张;热响应材料;

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