Biotechnology Letters

, Volume 36, Issue 2, pp 391–396 | Cite as

Aligning cells in arbitrary directions on a membrane sheet using locally formed microwrinkles

  • Akira C. Saito
  • Tsubasa S. Matsui
  • Masaaki Sato
  • Shinji Deguchi
Original Research Paper
First Online:
Received:
Accepted:

Abstract

Sheets of cells can be used for tissue regenerative medicine. Cell alignment within the sheet is now a key factor in the next generation of this technology. Anisotropic cell sheets without random cell orientations have been conventionally produced with photolithographically, microfabricated substrates using special facilities and equipment. Here we demonstrate a more accessible approach to the fabrication of anisotropic substrates. We locally deformed part of an elastic membrane and simultaneously oxidized the surface to create microwrinkles as well as to enable adhesion to the extracellular matrix. The approach with the local loading made it possible to orient cells in controlled directions within a single membrane sheet depending on the strains determined by the controllable deformation. This technique potentially enables a versatile design of microwrinkles for target-compatible cell alignments.

Keywords

Cell orientation Cell sheets Contact guidance Microwrinkles Nanotopography Orientation of cells Regenerative medicine Tissue engineering 
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Notes

Acknowledgments

We thank Yuka Minegishi and Hitomi Onodera for their technical assistance and Masakazu Kawashita, Zhixia Li, and Naoko Matsui for their help in SEM observations. This study was supported in part by the JSPS KAKENHI Grants (Nos. 11J07552 to TSM; 20001007 to MS; and 24650248 and 24680049 to SD). Author contributions: Conceived and designed the experiments: ACS TSM SD. Performed the research: ACS TSM SD. Analyzed the data: ACS. Contributed reagents/materials/analysis tools: MS SD. Wrote the paper: SD.

Supplementary material

10529_2013_1368_MOESM1_ESM.tiff (905 kb)
Supplementary Fig. 1 Circumferential alignment of other cell types. The upper, middle, and lower panels show confluent bovine aortic smooth muscle cells (BASMCs) (F-actin and DNA), sparse BASMCs (GFP-actin), and sparse Swiss 3T3 fibroblasts (GFP-actin), respectively. These images were acquired at a position that corresponds to 12 o’clock of Fig. 4b within a microwrinkled sheet. The arrow indicates the direction of the microgrooves. These primary BASMCs were cultured in DMEM supplemented with 10 % fetal bovine serum and 1 % penicillin–streptomycin. Swiss 3T3 fibroblasts were cultured in high-glucose (4.5 g glucose l−1) DMEM supplemented with 10 % fetal bovine serum and 1 % penicillin–streptomycin. These cells were maintained in a 5 % CO2 incubator at 37 °C before the experiments and were imaged with the ORCA-R2 camera on the IX-71 microscope (TIFF 904 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Akira C. Saito
    • 1
  • Tsubasa S. Matsui
    • 2
  • Masaaki Sato
    • 1
    • 3
  • Shinji Deguchi
    • 3
    • 4
  1. 1.Department of Biomedical EngineeringTohoku UniversitySendaiJapan
  2. 2.Department of Biomolecular SciencesTohoku UniversitySendaiJapan
  3. 3.Department of Bioengineering and RoboticsTohoku UniversitySendaiJapan
  4. 4.Department of Nanopharmaceutical SciencesNagoya Institute of TechnologyNagoyaJapan

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