Ensemble stimulation of embryoid bodies using microfabricated ITO substrates.
Precise control of differentiation processes of pluripotent stem cell is key component for realization of regenerative medicine. Electrical stimulation is one of the promising techniques, particularly for regulation of neuronal regeneration. In the present study, we developed substrates with embedded electrodes, which allowed ensemble electrical stimulation of embryoid bodies (EBs) of stem cells. Microcavity-array patterns were fabricated on substrates with embedded electrodes using standard photo-lithography. Uniform-size EBs of P19 cells were prepared, inserted one by one in each microcavity, and electrical stimulation was applied through substrate electrodes. Stimulus-induced intracellular calcium transients were successfully monitored by fluorescence imaging. The results suggested that this method would be useful for applying precisely-controlled electrical stimulation to a large number of EBs of stem cells.
Takayama
Yuzo
Y
Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, University of Tokyo, Chiba, Japan. y-taka@bmpe.k.u-tokyo.ac.jp
Moriguchi
Hiroyuki
H
Saito
Atushi
A
Kotani
Kiyoshi
K
Jimbo
Yasuhiko
Y
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D004558:Electric Stimulation / Q000379:methods*; D047108:Embryonic Development / Q000502:physiology* / Q000528:radiation effects; D053595:Embryonic Stem Cells / Q000502:physiology* / Q000528:radiation effects; D051379:Mice; D008904:Miniaturization; D009474:Neurons / Q000502:physiology* / Q000528:radiation effects; D009924:Organ Culture Techniques / Q000379:methods*