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Three-Dimensional Cell Sheet Construction Method with a Polyester Micromesh Sheet.

Takeshi Hori1, Osamu Kurosawa1, Kohei Ishihara2

  • 1Compass to Healthy Life Research Complex Program, RIKEN, Kobe, Japan.

Tissue Engineering. Part C, Methods
|March 19, 2020
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Summary

This study introduces an improved micromesh culture system for creating robust, easy-to-handle three-dimensional (3D) cell sheets. This enhanced method facilitates widespread use in regenerative medicine and research, offering better cell observation and nutrient supply.

Keywords:
3Dcell sheetin vitromicromesh culturepolyester

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Regenerative Medicine

Background:

  • Cell sheet engineering is crucial for regenerative medicine and transplantation.
  • Previous micromesh cultures enabled cell sheet formation on microstructured mesh sheets.
  • Existing methods have limitations in robustness, handling, cost, and microscopic observation.

Purpose of the Study:

  • To develop a more usable micromesh culture system and devices for widespread application.
  • To enhance cell sheet construction for improved handling, robustness, and cost-effectiveness.
  • To enable controlled cell orientation and multilayered cell sheet formation.

Main Methods:

  • Utilized polyester micromesh sheets fixed in 3D-printed or polyoxymethylene (POM) frames.
  • Cultured various cells (fibroblasts, hepatoma cells, mesenchymal stem cells) on the micromesh.
  • Employed optical coherence tomography and confocal microscopy for structural and cellular analysis.

Main Results:

  • Successfully formed robust, 100-400 μm thick cell sheets within 16 days.
  • Demonstrated mesh sheet integration within the cell sheet structure.
  • Showcased controlled cell alignment and the construction of multilayered cell sheets.
  • Achieved mass production of large cell sheets using POM devices.

Conclusions:

  • The enhanced micromesh culture system provides a robust, cost-effective, and user-friendly method for 3D cell sheet construction.
  • This technique offers advantages for cell observation, nutrient supply, and cell orientation control.
  • The system holds significant potential for future applications in medical and research fields.
  • The use of POM devices facilitates scalable production of cell sheets.