Gene Transfer Into the Donor Heart During Cold Preservation for Heart Transplantation Satoshi Gojo, Kazuo Niwaya, Shigeki Taniguchi, Kazuhiko Nishizaki,

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Mark I. Block, MD The Annals of Thoracic Surgery

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Gene Transfer Into the Donor Heart During Cold Preservation for Heart Transplantation Satoshi Gojo, Kazuo Niwaya, Shigeki Taniguchi, Kazuhiko Nishizaki, Soichiro Kitamura The Annals of Thoracic Surgery Volume 65, Issue 3, Pages 647-652 (March 1998) DOI: 10.1016/S0003-4975(97)01295-2

Fig. 1 Macroscopic photograph of an ex vivo gene-transferred heart on postoperative day 7. In this photograph, cells positive for β-gal activity were recognized in the ventricles as blue spots. The bar indicates 1 mm of length. The Annals of Thoracic Surgery 1998 65, 647-652DOI: (10.1016/S0003-4975(97)01295-2)

Fig. 2 (A and B) X-gal and hematoxylin and eosin staining of donor hearts perfused with 1 × 109 pfu of the recombinant adenovirus Adex1CALacZ on postoperative day 7. Blue-stained cardiomyocytes are the cells expressing β-gal. Leukocyte infiltration is slightly detectable within the myocardium. (C) Histologic photograph of the adenovirus-negative control hearts. Blue cells are absent. (Magnification, ×200 before 35% reduction.) The Annals of Thoracic Surgery 1998 65, 647-652DOI: (10.1016/S0003-4975(97)01295-2)

Fig. 3 Microscopic photographs of hearts perfused with 1 × 1010 (A) and 1 × 1011 (B) pfu of adenovirus Adex1CALacZ 7 days after transplantation. Numerous leukocytes are present around the gene-transferred cells. (Magnification ×200.) On postoperative day 21, gene-transferred cells are absent within the myocardium. The perfusion of 1 × 1010 (C) and 1 × 1011 pfu (D) of adenovirus produced inflammatory reactions throughout the myocardium. (Magnification, ×100 before 35% reduction.) The Annals of Thoracic Surgery 1998 65, 647-652DOI: (10.1016/S0003-4975(97)01295-2)

Fig. 4 DNA prepared from the lung (lane 3), brain (lane 4), liver (lane 5), spleen (lane 6), kidney (lane 7), ovary (lane 8), and donor heart (lane 2). Lane 1 contains molecular weight markers (500–base pair [bp] ladder; top: 4 kilobase pairs, bottom: 500 bp). The primer pair used in this study detects the part (1,036 bp) of the Escherichia coli β-gal gene. From lane 3 to lane 8, the β-gal gene is not detectable. In lane 2, the band of β-gal is shown. This indicates that the heart was transfected successfully by the recombinant adenovirus. The Annals of Thoracic Surgery 1998 65, 647-652DOI: (10.1016/S0003-4975(97)01295-2)