title: New model for cardiomyocyte sheet transplantation using a virus-cell fusion technique.
abstract: OBJECTIVE
  To facilitate close contacts between transplanted cardiomyocytes and host skeletal muscle using cell fusion mediated by hemagglutinating virus of Japan envelope (HVJ-E) and tissue maceration.
  
  
  METHODS
  Cardiomyocytes (1.5 × 10(6)) from fetal rats were first cultured. After proliferation, some cells were used for fusion with adult muscle fibers using HVJ-E. Other cells were used to create cardiomyocyte sheets (area: about 3.5 cm(2) including 2.1 × 10(6) cells), which were then treated with Nile blue, separated, and transplanted between the latissimus dorsi and intercostal muscles of adult rats with four combinations of HVJ-E and/or NaOH maceration: G1: HVJ-E(+), NaOH(+), Cardiomyocytes(+); G2: HVJ-E(-), NaOH(+), Cardiomyocytes(+); G3: HVJ-E(+), NaOH(-), Cardiomyocytes(+); G4: HVJ-E(-), NaOH(-), Cardiomyocytes(-). At 1 and 2 wk after transplantation, the four groups were compared by detection of beating domains, motion images using moving target analysis software, action potentials, gene expression of MLC-2v and Mesp1 by reverse transcription-polymerase chain reaction, hematoxylin-eosin staining, and immunostaining for cardiac troponin and skeletal myosin.
  
  
  RESULTS
  In vitro cardiomyocytes were fused with skeletal muscle fibers using HVJ-E. Cardiomyocyte sheets remained in the primary transplanted sites for 2 wk. Although beating domains were detected in G1, G2, and G3 rats, G1 rats prevailed in the number, size, motion image amplitudes, and action potential compared with G2 and G3 rats. Close contacts were only found in G1 rats. At 1 wk after transplantation, the cardiomyocyte sheets showed adhesion at various points to the myoblast layer in the latissimus dorsi muscle. At 2 wk after transplantation, close contacts were seen over a broad area. Part of the skeletal muscle sarcoplasma seemed to project into the myocardiocyte plasma and some nuclei appeared to share both sarcoplasmas.
  
  
  CONCLUSIONS
  The present results show that close contacts were acquired and facilitated the beating function, thereby providing a new cellular transplantation method using HVJ-E and NaOH maceration.
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Yuto
authors.initials: Y
authors.lastname: Takahashi
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Daihachiro
authors.initials: D
authors.lastname: Tomotsune
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Sakiko
authors.initials: S
authors.lastname: Takizawa
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Fengming
authors.initials: F
authors.lastname: Yue
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Nagai
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yokoyama
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Kanji
authors.initials: K
authors.lastname: Hirashima
authors.affiliation: Yuto Takahashi, Daihachiro Tomotsune, Fengming Yue, Kanji Hirashima, Katsunori Sasaki, Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Sasaki
keywords: Cardiomyocyte sheet; Cell fusion; Cellular transplantation method; Hemagglutinating virus of Japan envelope; Latissimus dorsi; Maceration; NaOH