Improved heart function with myogenesis and angiogenesis after autologous porcine bone marrow stromal cell transplantation Shinji Tomita, MD, PhD, Donald.

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Improved heart function with myogenesis and angiogenesis after autologous porcine bone marrow stromal cell transplantation Shinji Tomita, MD, PhD, Donald A.G. Mickle, MD, Richard D. Weisel, MD, Zhi-Qiang Jia, MD, Laura C. Tumiati, BSc, Yasmin Allidina, RTNM, Peter Liu, MD, Ren-Ke Li, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 123, Issue 6, Pages 1132-1140 (June 2002) DOI: 10.1067/mtc.2002.120716 Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Appearance of 4-week-old 5-azacytidine-treated bone marrow stromal cells in culture immediately before transplantation (original magnification 400×). Staining was positive for vimentin (stromal cell marker, b), α-smooth muscle actin (c), and desmin (myogenic progenitor marker, d) in 100%, 80% ± 6%, and 30% ± 8% of cells, respectively. Large arrows indicate positively stained cells and small arrows indicate the negatively stained cells). When the induced stromal cell culture became confluent, myocyte was present (a, arrow). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Tissue slices obtained 4 weeks after implantation from transplanted area stained for BrdU (a) and troponin I (b). Islands of cardiac tissue in scar area of transplanted hearts stained positively for both BrdU and troponin I (original magnification 400×). a, Islands contained BrdU-stained differentiated cardiomyocytes (small arrows) with organized sarcomeres and Z-bands; some transplanted cells (BrdU-positive cells) also differentiated into interstitial cells in fibrotic tissue (open arrows; original magnification 400×). b, Large arrows point to intercalated discs (original magnification 400×). c, Hematoxylin and eosin-stained tissue showing transplanted muscle (m; original magnification 40×). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Immunohistochemical staining for BrdU of myocardial scar tissue 4 weeks after cell transplantation (original magnification 100×). Some endothelial cells in blood vessels in transplanted region stained positively for BrdU (arrow). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Changes in resting regional perfusion (a), wall motion (b), and wall thickness (c) of myocardial infarct region in transplant (n = 5) and control (n = 6) groups from before transplantation to 4 weeks after transplantation, as measured by the MIBI scan. Regional perfusion in transplant group was maintained, whereas that in control groups decreased (P <.05). Regional wall motion and wall thickness were increased (P <.05) in transplant group. Bar heights represent mean; error bars represent SE. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Representative 3-dimensional views of reconstructed hearts from gated MIBI scan. Animals in control (a) and transplant (b) groups were scanned at 4 weeks after cell transplantation. a, In control group, end-diastolic volume was 82 mL, end-systolic volume was 52 mL, and ejection fraction was 37%. b, In transplant group, end-diastolic volume was 72 mL, end-systolic volume was 40 mL, and ejection fraction was 50%. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 6 Differences in stroke volumes (a) and ejection fractions (b) from before transplantation to 4 weeks after transplantation, as measured by MIBI scan, in transplant (n = 5) and control (n = 6) groups. Stroke volume in transplant group was greater (P <.05) than in control group. Although difference in ejection fraction appeared increased relative to that in control hearts, differences did not differ statistically significantly between groups (NS). Bar heights represent mean; error bars represent SE. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 7 Heart function was measured with Millar and conductance catheters (measuring pressure and volume) at 4 weeks after cell transplantation, and left ventricular stroke work and end-diastolic volume relations are shown. Slope of preload-recruitable stroke work in transplant group (diamonds) was greater (P <.05) than that in control group (circles). Data points represent mean; error bars represent SE. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1132-1140DOI: (10.1067/mtc.2002.120716) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions