Measuring Mechanical Properties of Cell Sheets by a Tensile Test Using a Self-Attachable Fixture
Kaoru Uesugi*, Yoshitake Akiyama*, Takayuki Hoshino*,
Yoshikatsu Akiyama**, Masayuki Yamato**, Teruo Okano**,
and Keisuke Morishima*, †
*Department of Mechanical Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
**Institute of Advanced BioMedical Engineering and Science, Tokyo Women’s Medical University, 8-1 Kawadacho, Shinjuku, Tokyo 162-8666, Japan
In this paper, we present a tensile test system and a new self-attachable fixture for measuring mechanical properties of cell sheets. Previously when measuring the mechanical properties of cell sheets, the sheets would be damaged because they dried out. With our system, the tensile test can be carried out while the cell sheets are in the culture medium, so there is no damage by drying. Previously, it has also been difficult to attach a cell sheet in the tensile test system owing to the structure of the conventional fixture, and there has been no tensile test system which had a measurement range that covered the tension force range of the cell sheets. Therefore, we have addressed these problems by developing a self-attachable fixture and a tensile test system. To confirm suitability of the fixture and test system, we measured mechanical properties of two different kinds of cultured cell sheets, C2C12 (cells mouse myoblast cells) and NIH-3T3 (3T3) cells (mouse fibroblast cells), and of the same kinds of sheets treated with cytochalasin-D. We confirmed differences in mechanical properties for each kind of cell sheet. This indicates that our new fixture and test system are applicable for measurement of mechanical properties of cell sheets without damage of the sheets by drying.
Yoshikatsu Akiyama, Masayuki Yamato, Teruo Okano, and
and Keisuke Morishima, “Measuring Mechanical Properties of Cell Sheets by a Tensile Test Using a Self-Attachable Fixture,” J. Robot. Mechatron., Vol.25, No.4, pp. 603-610, 2013.
-  M. Yamato and T. Okano, “Cell sheet engineering,” Materials Today, Vol.7, No.5, pp. 42-47, 2004.
-  I. E. Hannachi, M. Yamato, and T. Okano, “Cell sheet technology and cell patterning for biofabrication,” Biofabrication, Vol.1, No.2, pp. 1-13, 2009.
-  I. E. Hannachi, M. Yamato, and T. Okano, “Cell sheet engineering: a unique nanotechnology for scaffold-free tissue reconstruction with clinical applications in regenerative medicine,” J. Internal Med., Vol.267, No.1, pp. 54-70, 2010.
-  T. Shimizu, M. Yamato, A. Kikuchi, and T. Okano, “Cell sheet engineering for myocardial tissue reconstruction,” Biomaterials, Vol.24, No.13, pp. 2309-2316, 2003.
-  R. Uchida, N. Tanaka, M. Higashimori, K. Tadakuma, M. Kaneko, M. Kondo, and M. Yamato, “Cell sheet stiffness sensing without taking out from culture liquid,” Conf Proc. IEEE Eng. Med. Biol. Soc., pp. 827-830, 2010.
-  R. F. Ker, “Soft composites,” in J. F. V. Vincent (Ed.), Biomechanics-Materials: A Practical Approach, Oxford University Press, Ch. 7, 1992.
-  T. Matsumoto, J. Sato, M. Yamamoto, and M. Sato, “Smooth muscle cells freshly isolated from rat thoracic aortas are much stiffer than cultured bovine cells: Possible effect of phenotype,” JSME Int. J. Series C, Mechanical Systems, Machine Elements and Manufacturing, Vol.43, No.4, pp. 867-874, 2000.
-  K. Nagayama, Y. Nagano, M. Sato, and T. Matsumoto, “Effect of actin filament distribution on tensile properties of smooth muscle cells obtained from rat thoracic aortas,” J. Biomech., Vol.39, No.2, pp. 293-301, 2006.
-  J. S. Rossmann, “Elastomechanical properties of bovine veins,” J. Mech. Behav. Biomed. Mater, Vol.3, No.2, pp. 210-215, 2010.
-  V. Kanyanta and A. Ivankovic, “Mechanical characterisation of polyurethane elastomer for biomedical applications,” J. Mech. Behav. Biomed. Mater, Vol.3, No.1, pp. 51-62, 2010.
-  A. P. Jackson, “Bone, nacre, and other ceramics,” in J. F. V. Vincent (Ed.), Biomechanics-Materials: A Practical Approach Oxford University Press, Ch. 3, 1992.
-  C. Bellini, P. Glass, M. Sitti, and E. S. Di Martino, “Biaxial mechanical modeling of the small intestine,” J. Mech. Behav. Biomed. Mater, Vol.4, No.8, pp. 1727-1740, 2011.
-  Y. Lanir and Y. C. Fung, “Two-dimensional mechanical properties of rabbit skin-II. Experimental results,” J. Biomech., Vol.7, No.2, pp. 171-182, 1974.
-  Y. Tsuda, A. Kikuchi, M. Yamato, A. Nakao, Y. Sakurai, M. Umezu, and T. Okano, “The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets,” Biomaterials, Vol.26, No.14, pp. 1885-1893, 2005.
-  K. Uesugi, Y. Akiyama, T. Hoshino, Y. Akiyama, M. Yamato, T. Okano, and K. Morishima, “Measuring adhesion force of a cell sheet by the ninety-degree peel test using a multi hook type fixture,” J. of Biomechanical Science and Engineering, Vol.8, No.2, pp. 129-138, 2013.
-  N. Yamada, T. Okano , H. Sakai, F. Karikusa, Y. Sawasaki, and Y. Sakurai, “Thermo-responsive polymeric surfaces; control of attachment and detachment of cultured cells,” Macromol Rapid Commun, Vol.11, pp. 571-576, 1990.
-  V. Lulevich, T. Zink, H. Y. Chen, F. T. Liu, and G. Liu, “Cell mechanics using atomic force microscopy-based single-cell compression,” Langmuir, Vol.22, No.19, pp. 8151-8155, 2006.
-  J. L. Tan, J. Tien, D. M. Pirone, D. S. Gray, K. Bhadriraju, and C. S. Chen, “Cells lying on a bed of microneedles: an approach to isolate mechanical force,” Proc. Natl. Acad. Sci. USA, Vol.100, No.4, pp. 1484-1489, 2003.
-  K. Nagayama and T. Matsumoto, “Contribution of actin filaments and microtubules to quasi-in situ tensile properties and internal force balance of cultured smooth muscle cells on a substrate,” Am. J. Physiol. Cell Physiol., Vol.295, No.6, pp. C1569-1578, 2008.
-  Y. Haraguchi, T. Shimizu, T. Sasagawa, H. Sekine, K. Sakaguchi, T. Kikuchi, W. Sekine, S. Sekiya, M. Yamato, M. Umezu, and T. Okano, “Fabrication of functional three-dimensional tissues by stacking cell sheets in vitro,” Nat. Protoc., Vol.7, No.5, pp. 850-858, 2012.
-  T. Matsumoto and K. Nagayama, “Tensile properties of vascular smooth muscle cells: bridging vascular and cellular biomechanics,” J. Biomech., Vol.45, No.5, pp. 745-755, 2012.
Copyright© 2013 by Fuji Technology Press Ltd. and Japan Society of Mechanical Engineers. All right reserved.