title: Simultaneous induction of calcium transients in embryoid bodies using microfabricated electrode substrates.
abstract: Precise control of differentiation processes of pluripotent stem cells is a key component for the further development of regenerative medicine. For this purpose, combining a cell-aggregate-size treatment for regulating intercellular signal transmissions and an electrical stimulation technique for inducing cellular responses is a promising approach. In the present study, we developed microfabricated electrode substrates that allow simultaneous stimulation of embryoid bodies (EBs) of P19 cells. Mouse embryonal carcinoma P19 cells can be induced to differentiate into three germ layers and serve as a promising stem cell model. Microcavity-array patterns were fabricated onto indium-tin-oxide (ITO) substrates using a standard photo-lithography technique, and uniform-sized EBs of P19 cells were inserted into each microcavity. Electrical stimulation was applied to the EBs through substrate electrodes and stimulus-induced intracellular calcium transients were monitored. We confirmed that the developed electrode device could simultaneously stimulate smaller (200μm diameter) and larger (500μm diameter) EBs inserted in the microcavities and induce specific spatio-temporal patterns of intracellular calcium transients in the EBs with fine reproducibility. We concluded that the developed microcavity array with embedded electrodes could simultaneously and effectively stimulate uniform-sized EBs inserted in it. Therefore, it is a promising experimental tool for precisely controlling cell differentiation processes.
authors: Graduate School of Information Science and Technology, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. yuzo_takayama@ipc.i.u-tokyo.ac.jp
authors: Yuzo
authors: Y
authors: Takayama
authors: Atsushi
authors: A
authors: Saito
authors: Hiroyuki
authors: H
authors: Moriguchi
authors: Kiyoshi
authors: K
authors: Kotani
authors: Takafumi
authors: T
authors: Suzuki
authors: Kunihiko
authors: K
authors: Mabuchi
authors: Yasuhiko
authors: Y
authors: Jimbo
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D048429:Cell Size; D004558:Electric Stimulation; D004566:Electrodes; D058732:Embryoid Bodies; D005855:Germ Layers / Q000166:cytology; D051379:Mice; D055616:Microtechnology; D039904:Pluripotent Stem Cells / Q000166:cytology*; D015203:Reproducibility of Results
chemical_list: D002118:Calcium