Biomedical Microdevices

, Volume 11, Issue 2, pp 413–419

Localized electrical stimulation to C2C12 myotubes cultured on a porous membrane-based substrate

  • Takeshi Ishibashi
  • Yu Hoshino
  • Hirokazu Kaji
  • Makoto Kanzaki
  • Masaaki Sato
  • Matsuhiko Nishizawa
Article

DOI: 10.1007/s10544-008-9247-7

Cite this article as:
Ishibashi, T., Hoshino, Y., Kaji, H. et al. Biomed Microdevices (2009) 11: 413. doi:10.1007/s10544-008-9247-7

Abstract

We report a porous membrane-based cell culture device that can conduct localized electrical stimulation of a cell monolayer. The device’s cell culture substrate is a microporous alumina membrane with an underlying thin poly(dimethylsiloxane) (PDMS) film spotted with holes. When electric current is generated between the device’s Pt ring electrodes—one of which is placed above the cells and the other below the PDMS layer—the current density condenses at the holes in the PDMS film, and cells located above the holes can be electrically stimulated. C2C12 cells were confluently cultured on the substrate and were differentiated to myotubes. To control the stimulated area in the substrate, we attempted to seal and reopen the holes of the PDMS film by using an air bubble. Since the current pulse could be effectively blocked at the sealed holes, fluorescent Ca2+ transients, indicative of cellular excitation, were observed from the myotubes located above holes in the open state.

Keywords

Electrical stimulation C2C12 myotube Ca2+ transient Porous alumina membrane Bubble-sealing 

Supplementary material

10544_2008_9247_MOESM1_ESM.doc (248 kb)
Suppl.Fig. 1 (a) Fluorescence image of Flou-4-loaded myotubes above the opened hole. Scale bar is 200 μm. (b) Relative intensity of Ca2+ transients measured at the point 1 denoted in (a). The fluorescence intensities of the first five transients at each given current were averaged and normalized to the value at the current amplitude of 1 mA. (DOC 247 KB)
10544_2008_9247_MOESM2_ESM.mpg (1.2 mb)
Suppl. Movie 1Ca2+ responses in myotubes in and near the opened hole during the application of the current pulses. (MPG 1.19 MB)
10544_2008_9247_MOESM3_ESM.mpg (1.9 mb)
Suppl. Movie 2Externally induced contraction of myotubes on the porous membrane substrate. (MPG 1.87 MB)

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Takeshi Ishibashi
    • 1
  • Yu Hoshino
    • 1
  • Hirokazu Kaji
    • 1
    • 4
  • Makoto Kanzaki
    • 2
    • 4
  • Masaaki Sato
    • 1
    • 3
  • Matsuhiko Nishizawa
    • 1
    • 4
  1. 1.Department of Bioengineering and Robotics, Graduate School of EngineeringTohoku UniversitySendaiJapan
  2. 2.Department of Biomedical Engineering, Graduate School of Biomedical EngineeringTohoku UniversitySendaiJapan
  3. 3.Department of Biomedical Engineering, Graduate School of Biomedical EngineeringTohoku UniversitySendaiJapan
  4. 4.JSTCRESTTokyoJapan

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