Tetsuro Sakai, MD, Ren-Ke Li, MD, MSc, PhD, Richard D

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The fate of a tissue-engineered cardiac graft in the right ventricular outflow tract of the rat Tetsuro Sakai, MD, Ren-Ke Li, MD, MSc, PhD, Richard D. Weisel, MD, Donald A.G. Mickle, MD, Eung T.Joong Kim, MD, PhD, Zhi-Qian Jia, MD, MSc, Terrence M. Yau, MD The Journal of Thoracic and Cardiovascular Surgery Volume 121, Issue 5, Pages 932-942 (May 2001) DOI: 10.1067/mtc.2001.113600 Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Adult heart cell-seeded patch in vitro. Hematoxylin and eosin staining of vertical sections after culturing for 3 weeks (A, 100×; B, 400×). The upper and lower surfaces were seeded sequentially. Full-thickness penetration of the cultured cells was found. The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Cell growth in the gelatin patch. A, Fetal cardiomyocyte (FCM). B, Adult cardiomyocyte (ACM). Each point represents the mean ± the standard deviation. The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Gross appearance of control patch at 4 weeks. A, The anterior view. B, Vertically sliced view. C, The endocardial surface view of the heart from a control rat. Note full thickness replacement of RVOT with the patch (arrows). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Thickness of the RVOT. Data are shown as the mean ± the standard deviation. The patches became significantly thinner with time as the scaffold dissolved in all groups. No difference in the thickness was found between the patch groups at 4 or 12 weeks. Compared with normal hearts, the fetal cardiomyocyte group was thicker (†P =.036) at 4 weeks, and the control group was thinner (‡P =.047) at 12 weeks. FCM, Fetal cardiomyocyte patch; ACM, adult cardiomyocyte patch. *P =.003. **P =.001. The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Histologic studies of RVOT patch. A fetal cardiomyocyte patch 1 day after the implantation had tissue formation (A, 100×; B, 400×). At 4 weeks an inflammatory reaction was seen in all groups as the gelatin scaffold dissolved. The adult group is shown in C (100×) and D (400×). The control group is shown in E (100×) and F (400×). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 6 Twelve weeks after the replacement (A, control group; B, adult group; C, fetal group; all magnified 100×). The gelatin structure of the patches had completely disappeared and the inflammatory reaction had resolved. Normal RVOT was shown for comparison (D, 100×). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 7 BrdU staining (B and D ) with the corresponding hematoxylin and eosin staining (A and C ) of the seeded patches at 12 weeks after the replacement (A and B, fetal group; C and D, adult group; 400×). Some of the cells in the patched area stained positively (brown) for BrdU, indicating that they contained cultured cells seeded in the gelatin mesh. The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 8 Factor VIII staining (dark red) identified endothelial cells on the surface of the patch at 4 weeks after implantation in all groups. The fetal cardiomyocyte group is illustrated in A (400×) and the normal endocardial surface is shown in B (400×). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 932-942DOI: (10.1067/mtc.2001.113600) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions