Abstract
We have fabricated a simple Si-MEMS device consisting of a microcantilever and a base to measure active tension generated by skeletal muscle myotubes derived from murine myoblast cell line C2C12. We have developed a fabrication process for integration of myotubes onto the device. To position myotubes over the gap between the cantilever and the base without damage due to mechanical peeling or the use of an enzymatic reaction, we cultured myotubes on poly-N-isopropylacrylamide (PNIPAAm) as a sacrifice layer. By means of immune staining of α-actinin, it was confirmed that a myotube micropatterned onto the device bridged the gap between the cantilever and the base. After 7d differentiation, the myotube was actuated by electrical stimulation. The active tension generated by the myotube was evaluated by measuring the bending of the cantilever using image processing. On twitch stimulation, the myotube on the device contracted and generated active tension in response to the electrical signals. On tetanus tension measurement, approximately 1.0 μN per single myotube was obtained. The device developed here can be used in wide area of in vitro skeletal muscle studies, such as drug screening, physiology, regenerative medicine, etc.
Similar content being viewed by others
References
R.G. Dennis, P.E. Kosnik 2nd, M.E. Gilbert, J.A. Faulkner, Am. J. Physiol. Cell. Physiol. 280, C288 (2001)
A.W. Feinberg, A. Feigel, S.S. Shevkoplyas, S. Sheehy, G.M. Whitesides, K.K. Parker, Science 317, 1366 (2007)
H. Fujita, T. Nedachi, M. Kanzaki, Exp. Cell Res. 313, 1853 (2007)
H. Fujita, K. Shimizu, E. Nagamori, Biotechnol. Bioeng. 103, 1034 (2009)
T.J. Hawke, D.J. Garry, J. Appl. Physiol. 91, 534 (2001)
Y. Hiratsuka, M. Miyata, T. Tada, T.Q. Uyeda, Proc. Natl. Acad. Sci. U. S. A. 103, 13618 (2006)
J. Kim, J. Park, S. Yang, J. Baek, B. Kim, S.H. Lee et al., Lab. Chip. 7, 1504 (2007)
G. Lin, R.E. Palmer, K.S. Pister, K.P. Roos, IEEE Trans. Biomed. Eng. 48, 996 (2001)
D.K. McMahon, P.A.W. Anderson, R. Nassar, J.B. Bunting, Z. Saba, A.E. Oakeley et al., Am. J. Physiol. Cell. Physiol. 35, c1795 (1994)
S. Nishimura, A. Yamada, M. Katoh, K.P. Yamada, H. Yamashita, Y. Saeki et al., Am. J. Physiol. Heart. Circ. Physiol. 287, H196 (2004)
H. Park, R. Bhalla, R. Saigal, M. Radisic, N. Watson, R. Langer et al., J. Tissue. Eng. Regen. Med. 2, 279 (2008)
P.R. Shepherd, B.B. Kahn, N. Engl. J. Med. 341, 248 (1999)
K. Shimizu, H. Fujita, E. Nagamori, J. Biosci. Bioeng. (2009). doi: 10.1016/j.jbiosc.2009.07.016
R.K. Soong, G.D. Bachand, H.P. Neves, A.G. Olkhovets, H.G. Craighead, C.D. Montemagno, Science 290, 1555 (2000)
Y. Tanaka, K. Morishima, T. Shimizu, A. Kikuchi, M. Yamayo, T. Okano et al., Lab. Chip 6, 362 (2006)
H. Vandenburgh, J. Shansky, F. Benesch-Lee, V. Barbata, J. Reid, L. Thorrez et al., Muscle Nerve 37, 438 (2008)
K. Wilson, P. Molnar, J.J. Hickman, Lab. Chip 7, 920 (2007)
J. Xi, J.J. Schmidt, C.D. Montemagno, Nat. Mater. 4, 180 (2005)
D. Yaffe, O. Saxel, Nature 270, 725 (1977)
Acknowledgements
The authors would like to thank Yuki Morioka for the help with the cell culture.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplementary movie 1
Movement of the myotube and the microcantilever on 1 Hz electric stimulation (M1V 1.37 MB)
Supplementary movie 2
Movement of the myotube and the microcantilever under tetanus condition (20 Hz) (M1V 1405 kb) (M1V 1.23 MB)
Rights and permissions
About this article
Cite this article
Shimizu, K., Sasaki, H., Hida, H. et al. Assembly of skeletal muscle cells on a Si-MEMS device and their generative force measurement. Biomed Microdevices 12, 247–252 (2010). https://doi.org/10.1007/s10544-009-9379-4
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10544-009-9379-4