Assembly of skeletal muscle cells on a Si-MEMS device and their generative force measurement.
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 alpha-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 microN 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.
Shimizu
Kazunori
K
Toyota Central R&D Labs Inc, 41-1 Yokomichi, Nagakute, Aichi 480-1192, Japan.
Sasaki
Hikaru
H
Hida
Hirotaka
H
Fujita
Hideaki
H
Obinata
Kei
K
Shikida
Mitsuhiro
M
Nagamori
Eiji
E
D000178:Acrylamides / Q000737:chemistry; D000180:Acrylic Resins / Q000737:chemistry; D000185:Actinin / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000502:physiology; D018482:Muscle, Skeletal / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D055585:Physical Phenomena
D000178:Acrylamides; D000180:Acrylic Resins; D000185:Actinin; C052970:poly-N-isopropylacrylamide; C067295:N-isopropylacrylamide