Development of abnormal tissue architecture in transplanted neonatal rat myocytes Peter Whittaker, PhD, Jochen Müller-Ehmsen, MD, Joan S Dow, BS, Larry.

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Development of abnormal tissue architecture in transplanted neonatal rat myocytes Peter Whittaker, PhD, Jochen Müller-Ehmsen, MD, Joan S Dow, BS, Larry H Kedes, MD, Robert A Kloner, MD, PhD The Annals of Thoracic Surgery Volume 75, Issue 5, Pages 1450-1456 (May 2003) DOI: 10.1016/S0003-4975(02)04995-0 Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Micrographs of picrosirius red-stained sections viewed with circularly polarized light. Muscle cells appear green with cross-striations and collagen fibers appear green, yellow, or orange depending upon their thickness; myocardium distant from the infarct at 6 months (A), transplant tissue at (B) 2 weeks and (C) 6 months. In host tissue muscle cells are highly aligned, and the relatively small amount of collagen present is predominantly aligned parallel to the muscle. Although transplanted cells developed cross-striations after 2 weeks, they appeared small and disorganized. The collagen at the top and bottom of figure B is scar tissue associated with the injection. The transplant (central region) contains thinner collagen fibers. At 6 months, the cells and collagen fibers have increased in size; however, both lack the host tissue’s coherent organization. (Bar = 20 μm in all panels.) The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Retardation of transplant cells expressed as a percent of the host cells’ retardation. Although there was a progressive increase, transplant cell retardation never attained that of host cells. The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Average angular deviation of muscle cell orientation distributions obtained from transplant and mid-myocardial host tissue. Distributions from transplant cells had higher angular deviations than host cells (p < 0.05), an indicator of disarray. The 12 week group was excluded from statistical analysis because of the small sample size. The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Form factors for transplant and host cell nuclei. The transplant cells’ form factors were greater than that of host cells (p < 0.001), indicating round versus elongated profiles. The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Micrographs illustrating cell nuclear shape differences (hematoxylin & eosin staining). (A) Cells from a sham-operated heart reveal archetypal elongated nuclei (white arrows). (B and C) Cells 6 months after transplant into an infarct have rounded nuclei (white arrows). Fibroblasts (white arrowheads) are also present. (Bar = 20 μm in all panels.) The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Collagen content expressed as a percent of area analyzed. No value was calculated for the 0 to 1 day transplant period. Collagen content was always higher in the transplant than host tissue (p < 0.01; 12 week group excluded from statistical analysis). The Annals of Thoracic Surgery 2003 75, 1450-1456DOI: (10.1016/S0003-4975(02)04995-0) Copyright © 2003 The Society of Thoracic Surgeons Terms and Conditions