Measurement of Contractile Force of Myotube on Scaffold of Thin Film with Micro-Pattern-Markers

摘要

A scaffold of a transparent film with micro pattern markers hasbeen designed to measure the contractile force of myotubeunder electric stimulation in vitro. The scaffold is made of athin film (thickness 0.006 mm), of which the back side hasarrangement of polydimethylsiloxane micro-protrusions (0.004mm diameter, 0.002 mm height, 0.003 mm interval) made bythe photolithography technique. C2C12 (mouse myoblast) wasseeded on the film at the counter surface to the protrusions atthe density of 50000 cells/cm2. The cells were cultured on thescaffold for 12 days in the medium containing 10% FBS (fetalbovine serum) and 1% penicillin/ streptomycin at 310 K with5% of CO2 content. The electric pulses (amplitude of 30 V(0.06 A) ; pulse cycle of 1 s; pulse width of 1 ms) were appliedbetween electrodes of the titanium wire dipped in the medium.The contraction of myotubes is able to be observed through thetransparent scaffold at the microscope. The contractile force ofmyotube at the electric stimulation estimated by thedeformation of the film was 10－4 N. The designed scaffold hasa potential for the measurement of the local contractile force ofthe myotube microscopically in vitro.