Grafted skeletal myoblast sheets attenuate myocardial remodeling in pacing-induced canine heart failure model Hiroki Hata, MD, Goro Matsumiya, MD, PhD,

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Grafted skeletal myoblast sheets attenuate myocardial remodeling in pacing-induced canine heart failure model Hiroki Hata, MD, Goro Matsumiya, MD, PhD, Shigeru Miyagawa, MD, PhD, Haruhiko Kondoh, MD, PhD, Naomasa Kawaguchi, MD, PhD, Nariaki Matsuura, MD, PhD, Tatsuya Shimizu, MD, PhD, Teruo Okano, PhD, Hikaru Matsuda, MD, PhD, Yoshiki Sawa, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 132, Issue 4, Pages 918-924 (October 2006) DOI: 10.1016/j.jtcvs.2006.01.024 Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Manufacture and grafting of skeletal myoblast sheets. A, Primary cultures of autologous skeletal myoblasts showing a spindle-shaped appearance. B, Each graft was soft and limp, with a diameter of 30 to 40 mm. C, Cross-section of a graft, which was approximately 90-μm thick. D, Graft (arrows) was placed and smoothed out on the LV wall. Scale bar: 50 μm (A, C) and 10 mm (B). The Journal of Thoracic and Cardiovascular Surgery 2006 132, 918-924DOI: (10.1016/j.jtcvs.2006.01.024) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Echocardiographic findings. LVEF (A), LV%FS (B), and LVAWTh (C) increased, whereas LVESA (D) decreased after implantation in the Tx group. Data are expressed as the means ± standard error of mean. *P < .05 versus the C group. #P < .05 versus the baseline. The Journal of Thoracic and Cardiovascular Surgery 2006 132, 918-924DOI: (10.1016/j.jtcvs.2006.01.024) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Postsurgical histologic findings. Macroscopic findings of cross-sectional specimens of canine heart. Tx group (A) showed an increased LV wall thickness and decreased LV thickness, compared with C group (B). Photomicrograph of a section through the ventricular wall that was stained with hematoxylin-eosin and contained grafted cells that survived and formed layers over the epicardium (C). Higher magnification showing elongated skeletal myoblasts (arrows) with infiltrated, round inflammatory cells (D). Survival of the grafted myoblasts was confirmed in adjacent sections by staining with skeletal muscle-specific nebulin (E). Picrosirius red staining showed a significant reduction of fibrosis in Tx group (F) compared with C group (G). PCNA-positive cells were found in some areas of the myocardium in Tx group (arrows) (H), but none were identified in the C group (I). Higher magnification of PCNA-positive cells including cardiomyocyte with striated pattern (arrows) and interstitial cells (arrowhead) (J). The number of TUNEL-positive myocytes (arrows) was significantly lower in Tx group (K) than in C group (L). Scale bar: 20 mm (A, B); 100 μm (C, F-I, K, L); 40 μm (D, E); and 20 μm (J). The Journal of Thoracic and Cardiovascular Surgery 2006 132, 918-924DOI: (10.1016/j.jtcvs.2006.01.024) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions