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Control of Cell Adhesion using Hydrogel Patterning Techniques for Applications in Traction Force Microscopy
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Analysis of force vector field during centrifugation for optimizing cell sheet adhesion.

Yuji Haraguchi1, Yuki Kagawa2, Hirotsugu Kubo2

  • 1Institute of Advanced Biomedical Engineering and Science, TWIns, Tokyo Women's Medical University, Shinjuku-ku, Tokyo, Japan.

Biotechnology Progress
|June 1, 2019
PubMed
Summary

Centrifugation speeds up tissue fabrication by improving cell sheet adhesion. By stabilizing the centrifuge plate, researchers eliminated unwanted forces, reducing fabrication time from 5 minutes to 1 minute without cytotoxicity.

Keywords:
3D tissue engineeringacceleration vectorattitude angledeformationrapid fabricationsliding

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Area of Science:

  • Biomaterials Engineering
  • Tissue Engineering
  • Cell Biology

Background:

  • Centrifugation is utilized for fabricating three-dimensional tissues via cell sheet layering.
  • Optimal centrifugal force enhances cell sheet adhesion to culture dishes and between layers, reducing fabrication time.
  • High rotational speeds can induce cell sheet sliding and deformation, hindering further time reduction.

Purpose of the Study:

  • To investigate the theoretical basis of centrifugal force application during tissue fabrication.
  • To identify the cause of cell sheet sliding and deformation at high rotational speeds.
  • To optimize centrifugation protocols for faster and more efficient tissue engineering.

Main Methods:

  • Theoretical derivation of the spatial distribution of acceleration on a centrifuge plate.
  • Analysis of the force vector field acting on cell sheets.
  • Experimental manipulation of plate orientation using added weights to maintain horizontal positioning.
  • Evaluation of cell sheet adhesion, deformation, and cytotoxicity under optimized centrifugation.

Main Results:

  • The study theoretically derived the acceleration distribution on a centrifuge plate during rotation.
  • Negative effects like cell sheet sliding and deformation were attributed to the centrifugal force component parallel to the plate surface, caused by plate inclination.
  • Adding weights to the plate edge to maintain horizontal orientation eliminated these negative effects.
  • Optimized centrifugation significantly reduced the adhesion time for mouse myoblast sheets from 5 minutes to 1 minute without observable cytotoxicity.

Conclusions:

  • Maintaining a horizontal plate surface during centrifugation is crucial for preventing unwanted forces on cell sheets.
  • Optimized centrifugation protocols, by controlling force vectors, can drastically reduce tissue fabrication time.
  • This method offers a more efficient approach to tissue engineering, enhancing cell sheet adhesion and integrity.