ハラグチ ユウジ   Haraguchi Yuji
  原口 裕次
   所属   研究施設 研究施設
   職種   特任講師
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 Electrical coupling of cardiomyocyte sheets occurs rapidly via functional gap junction formation.
掲載誌名 正式名:Biomaterials
略  称:Biomaterials
ISSNコード:(0142-9612)0142-9612(Linking)
掲載区分国外
巻・号・頁 27(27),4765-74頁
著者・共著者 Haraguchi Yuji†, Shimizu Tatsuya, Yamato Masayuki, Kikuchi Akihiko, Okano Teruo*
発行年月 2006/09
概要 Previously, we have successfully created pulsatile myocardial tissue grafts using our novel technology, "cell sheet engineering", that layers cell sheets fabricated on temperature-responsive culture dishes to form three-dimensional (3-D) structures. Electrical coupling is established between layered neonatal rat cardiomyocyte sheets, resulting in the synchronized beating of 3-D myocardial tissues. However, the mechanism by which these layered cardiomyocyte sheets communicate electrically is not well-understood. In this study, we used a multiple-electrode extracellular recording system and demonstrated that bilayer cardiomyocyte sheets coupled electrically with slight delays 34+/-2 min (mean+/-SEM) after layering. These delays gradually decreased and the electrical actions of layered cell sheets were completely coupled 46+/-3 min (mean+/-SEM) after initial layering. Immunohistological analysis showed that connexin43, a gap junction (GJ)-related protein, existed not only at cell-to-cell interfaces but also on the free cell membrane in the cardiomyocyte sheet. Additionally, neither connexin40 nor connexin45, but only connexin43 was detected between bilayer cardiomyocyte sheets within 30 min after layering. Dye transfer assay demonstrated that the exchange of small molecules via GJs occurred within 30 min. The cell sheet manipulation technique using the temperature-responsive culture dishes has substantial advances and the exciting potential in the fields of cell and tissue physiology, as well as tissue engineering.
DOI 10.1016/j.biomaterials.2006.04.034
PMID 16737736