Advertisement

Quick and easy microfabrication of T-shaped cantilevers to generate arrays of microtissues

  • Benoît Kalman
  • Catherine Picart
  • Thomas BoudouEmail author
Article

Abstract

Over the past decade, a major effort was made to miniaturize engineered tissues, as to further improve the throughput of such approach. Most existing methods for generating microtissues thus rely on T-shaped cantilevers made by soft lithography and based on the use of negative SU-8 photoresist. However, photopatterning T-shaped microstructures with these negative photoresists is fastidious and time-consuming. Here we introduce a novel method to quickly generate T-shaped cantilevers dedicated to generation of cellular microtissues, based on the use of positive photoresist. With only two layers of photoresist and one photomask, we were able to fabricate arrays of microwells in less than 3 h, each containing two T-shaped cantilevers presenting either a rectangular or a circular geometry. As a proof of concept, these arrays were then replicated in poly(dimethylsiloxane) and microtissues composed of NIH 3T3 fibroblasts encapsulated in collagen I were generated, while the two cantilevers simultaneously constrain and report forces generated by the microtissues. Immunostainings showed longitudinally aligned and elongated fibroblasts over the whole microtissue after 8 days of culture. The method described here opens the potential to quick prototyping platforms for high-throughput, low-volume screening applications.

Keywords

Photolithography Positive photoresist PDMS Microtissues 

Notes

Acknowledgments

The authors thank the members of the technical staff of the PTA cleanroom in Grenoble for their technical support. This work was partly supported by the French RENATECH network. This work was supported by the European Commission, FP7 via an ERC Starting grant to C.P. (BIOMIM, GA 239370) and a PhD fellowship to BK.

References

  1. M. Ader, E. M. Tanaka, Curr. Opin. Cell Biol. 31, 23 (2014)CrossRefGoogle Scholar
  2. B. M. Baker, C. S. Chen, J. Cell Sci. 125, 3015 (2012)CrossRefGoogle Scholar
  3. T. Boudou, W. R. Legant, A. Mu, M. A. Borochin, N. Thavandiran, M. Radisic, P. W. Zandstra, J. A. Epstein, K. B. Margulies, C. S. Chen, Tissue Eng. A 18, 910 (2012)CrossRefGoogle Scholar
  4. F. Greiner, S. Quednau, F. Dassinger, R. Sarwar, H. F. Schlaak, M. Guttmann, P. Meyer, J. Micromech. Microeng. 23, 025018 (2013)CrossRefGoogle Scholar
  5. J. T. Hinson, A. Chopra, N. Nafissi, W. J. Polacheck, C. C. Benson, S. Swist, J. Gorham, L. Yang, S. Schafer, C. C. Sheng, A. Haghighi, J. Homsy, N. Hubner, G. Church, S. A. Cook, W. A. Linke, C. S. Chen, J. G. Seidman, C. E. Seidman, Science 349, 982 (2015)CrossRefGoogle Scholar
  6. B. Kalman, C. Monge, A. Bigot, V. Mouly, C. Picart, and T. Boudou. Comput. Methods Biomech. Biomed. Engin. 1 (2015).Google Scholar
  7. W. R. Legant, A. Pathak, M. T. Yang, V. S. Deshpande, R. M. McMeeking, C. S. Chen, Proc. Natl. Acad. Sci. U. S. A. 106, 10097 (2009)CrossRefGoogle Scholar
  8. J. Liu, D. Zhang, B. Sha, P. Yin, Z. Xu, C. Liu, L. Wang, F. Xu, L. Wang, Biomed. Microdevices 16, 655 (2014)CrossRefGoogle Scholar
  9. A. Ramade, W. R. Legant, C. Picart, C. S. Chen, T. Boudou, Methods Cell Biol. 121, 191 (2014)CrossRefGoogle Scholar
  10. M. S. Sakar, D. Neal, T. Boudou, M. A. Borochin, Y. Li, R. Weiss, R. D. Kamm, C. S. Chen, H. H. Asada, Lab Chip 12, 4976 (2012)CrossRefGoogle Scholar
  11. M. S. Sakar, J. Eyckmans, R. Pieters, D. Eberli, B. J. Nelson, C. S. Chen, Nat. Commun. 7, 11036 (2016)CrossRefGoogle Scholar
  12. H. Wang, A. A. Svoronos, T. Boudou, M. S. Sakar, J. Y. Schell, J. R. Morgan, C. S. Chen, V. B. Shenoy, Proc. Natl. Acad. Sci. U. S. A. 110, 20923 (2013)CrossRefGoogle Scholar
  13. A. R. West, N. Zaman, D. J. Cole, M. J. Walker, W. R. Legant, T. Boudou, C. S. Chen, J. T. Favreau, G. R. Gaudette, E. A. Cowley, G. N. Maksym, Am. J. Phys. Lung Cell. Mol. Phys. 304, L4 (2013)Google Scholar
  14. R. Zhao, T. Boudou, W.-G. Wang, C. S. Chen, D. H. Reich, Adv. Mater. 25, 1699 (2013)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Benoît Kalman
    • 1
    • 2
  • Catherine Picart
    • 1
    • 2
  • Thomas Boudou
    • 1
    • 2
    Email author
  1. 1.University Grenoble Alpes, LMGPGrenobleFrance
  2. 2.CNRS, LMGPGrenobleFrance

Personalised recommendations