Changfa Guo, MD, Husnain Kh

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Myoblast Transplantation for Cardiac Repair: From Automyoblast to Allomyoblast Transplantation Changfa Guo, MD, Husnain Kh. Haider, PhD, Chunsheng Wang, MD, Ru-San Tan, MBBS, Winston S.N. Shim, PhD, Philip Wong, MBBS, Eugene K.W. Sim, FRCS The Annals of Thoracic Surgery Volume 86, Issue 6, Pages 1841-1848 (December 2008) DOI: 10.1016/j.athoracsur.2008.08.038 Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Preparation of skeletal myoblasts (SkMs) and SkM survival after transplantation. (A) Green fluorescent cytoplasm showing desmin-positive SkMs (magnification ×600). (B) The SkMs were 74.96% pure (blue curve) for desmin expression when gated for control at 1.00% (red curve) by means of flow cytometric assay. (C) 4', 6'-diamidino-2-phenylindole hydrochloride (DAPI)–labeled SkMs with blue fluorescent nuclei (magnification ×200). (D) Merge of panel C and phase-contrast image to show total number of cells (magnification ×200). (E) Lac-z–labeled cells with bluish nuclei (magnification ×200). (F) The SkMs labeled with lac-z survived in the region of myocardial infarction on day 28 after transplantation (magnification ×12.5), and (G) with high magnification (×300). (H) The SkMs labeled with DAPI survived on day 28 (magnification ×150). The Annals of Thoracic Surgery 2008 86, 1841-1848DOI: (10.1016/j.athoracsur.2008.08.038) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Immunostaining and time courses of infiltration of macrophages (A and B), CD8+ cells (C and D), and CD4+ cells (E and F) into the cell injection site (magnification ×400). *Significantly increased numbers of infiltrating cells (including macrophages, CD4+, and CD8+ cells) as compared with control groups (p < 0.05). #Significantly increased numbers of infiltrating cells (including macrophages, CD4+, and CD8+ cells) in group 4 as compared with those in group 3 (p < 0.05). (Group 1 = dotted bars; group 2 = open bars; group 3 = brick bars; group 4 = solid bars.) The Annals of Thoracic Surgery 2008 86, 1841-1848DOI: (10.1016/j.athoracsur.2008.08.038) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Immunostaining and time courses of infiltration of macrophages (A and B), CD8+ cells (C and D), and CD4+ cells (E and F) into the cell injection site (magnification ×400). *Significantly increased numbers of infiltrating cells (including macrophages, CD4+, and CD8+ cells) as compared with control groups (p < 0.05). #Significantly increased numbers of infiltrating cells (including macrophages, CD4+, and CD8+ cells) in group 4 as compared with those in group 3 (p < 0.05). (Group 1 = dotted bars; group 2 = open bars; group 3 = brick bars; group 4 = solid bars.) The Annals of Thoracic Surgery 2008 86, 1841-1848DOI: (10.1016/j.athoracsur.2008.08.038) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Echocardiographic analysis showed the efficacy of skeletal myoblast (SkM) transplantation on cardiac function. The SkM transplantation improved cardiac ejection fraction (EF) (A) and fractional shortening (FS) (B), and limited the dilation of the left ventricle (C). *Significant improvement of EF/FS compared with baseline and those seen in control groups (p < 0.05). #Significant improvement of left ventricular end-diastolic dimension (LVEDD) as compared with baseline (p < 0.05). The Annals of Thoracic Surgery 2008 86, 1841-1848DOI: (10.1016/j.athoracsur.2008.08.038) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions