New model for cardiomyocyte sheet transplantation using avirus-cell fusion technique

• 作者: Yuto Takahashi, Daihachiro Tomotsune, Sakiko Takizawa, Fengming Yue, Mika Nagai, Tadayuki Yokoyama, Kanji Hirashima, and Katsunori Sasaki
• 来源期刊: 世界干细胞杂志：英文版（电子版）
• 学科: 医学
• 关键词: CARDIOMYOCYTE SHEET Latissimus dorsi Hemagglutinating virus of Japan ENVELOPE Cell fusion NAOH MACERATION Cellular TRANSPLANTATION method
• 年，卷（期）: 2015,（5）
• 页码范围: 883-893
• 页数: 11页
• 分类号: R

摘要

AIM To facilitate close contacts between transplantedcardiomyocytes and host skeletal muscle using cellfusion mediated by hemagglutinating virus of Japanenvelope （HVJ-E） and tissue maceration.METHODS： Cardiomyocytes （1.5 × 10^6） from fetalrats were first cultured. After proliferation, some cellswere used for fusion with adult muscle fibers usingHVJ-E. Other cells were used to create cardiomyocytesheets （area： about 3.5 cm^2 including 2.1 × 106cells）, which were then treated with Nile blue, separated,and transplanted between the latissimusdorsi and intercostal muscles of adult rats with fourcombinations 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 beatingdomains, motion images using moving target analysissoftware, action potentials, gene expression of MLC-2v and Mesp1 by reverse transcription-polymerasechain reaction, hematoxylin-eosin staining, and immunostainingfor cardiac troponin and skeletal myosin.RESULTS： In vitro cardiomyocytes were fused withskeletal muscle fibers using HVJ-E. Cardiomyocytesheets remained in the primary transplanted sites for2 wk. Although beating domains were detected inG1, G2, and G3 rats, G1 rats prevailed in the number,size, motion image amplitudes, and action potentialcompared with G2 and G3 rats. Close contacts wereonly found in G1 rats. At 1 wk after transplantation,the cardiomyocyte sheets showed adhesion at variouspoints to the myoblast layer in the latissimus dorsimuscle. At 2 wk after transplantation, close contactswere seen over a broad area. Part of the skeletalmuscle sarcoplasma seemed to project into themyocardiocyte plasma and some nuclei appeared toshare both sarcoplasmas.CONCLUSION： The present results show that closecontacts were acquired and facil