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J Thorac Cardiovasc Surg 1999;117:8-15
© 1999 Mosby, Inc.

CARDIOTHORACIC TRANSPLANTATION

LIPOSOME-MEDIATED GENE TRANSFER IN RAT LUNG TRANSPLANTATION: A COMPARISON BETWEEN THE IN VIVO AND EX VIVO APPROACHES

From the Division of Cardiothoracic Surgery,^a Department of Surgery, and Department of Pathology,^b Washington University School of Medicine, St Louis, Mo; Genzyme Corporation,^c Framingham, Mass; and Department of Surgery,^d University of Michigan, Ann Arbor, Mich.

*Supported by the Federal University of Rio de Janeiro–University Hospital Clementino Fraga, Filho, Brazil.

Read at the Seventy-eighth Annual Meeting of The American Association for Thoracic Surgery, Boston, Mass, May 3-6, 1998.

Received for publication May 8, 1998. Revisions requested June 15, 1998. Revisions received Aug 4, 1998. Accepted for publication Sept 1, 1998. Address for reprints: G. Alexander Patterson, MD, 3108 Queeny Tower, One Barnes-Jewish Hospital Plaza, St Louis, MO 63110.

Objective: We compared the efficacy of in vivo and ex vivo liposome transfection in rat lung transplantation.
Methods: (1) Chloramphenicol acetyltransferase group: Fischer rats underwent isogeneic transplantation (n = 4 per group). Recipients were put to death on postoperative day 2 for chloramphenicol acetyltransferase activity. Ex vivo setting: Grafts received cDNA complexed or not with liposomes and were transplanted after 1.5 or 10 hours at 10°C. In vivo setting: Donors were intravenously injected with cDNA complexed or not with liposomes. Lungs were harvested after 1.5 or 10 hours, preserved at 10°C, and transplanted. (2) Transforming growth factor–ß1 group: Brown-Norway rats served as donors and Fischer rats as recipients. All grafts were preserved for 3 hours at 10°C. On postoperative day 5, arterial oxygenation and histologic rejection scores were assessed. Ex vivo setting: Grafts received transforming growth factor–ß1 sense (n = 8) or antisense (n = 7) complexed with liposomes or cDNA alone (n = 5). In vivo setting: Donors were intravenously injected with liposome:transforming growth factor–ß1 sense cDNA (n = 7). Exposure time was 3 hours.
Results: (1) Chloramphenicol acetyltransferase–transfection was superior in the ex vivo group but was not statistically different for longer exposure times. (2) Transforming growth factor-ß1–arterial oxygenation was superior in the ex vivo liposome:sense group. cDNA alone was inefficient. Rejection scores were not statistically different between ex vivo and in vivo liposome:sense groups but were better when the ex vivo liposome:sense group was compared with the cDNA alone or the antisense groups.
Conclusions: (1) With current liposome technology, the ex vivo route is superior to the in vivo approach; (2) cDNA alone does not provide transgene expression at levels to produce a functional effect.

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