Optimization of delivery of adeno-associated virus mediated gene transfer to a transplanted heart in a rat model

Jiang Xiaofeng, Lars Burdorf, Rabea Hinkel, Christian Kupatt, Bruno Reichart, Ulrike Brandl, Michael Schmoeckel

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Objectives: To optimize transgene expression levels after Adeno-associated virus (AAV)-mediated gene transfer, different delivery methods were compared in a transplant setting. Materials and Methods: Heterotopic abdominal heart transplants were performed in male Lewis rats (250-280 g). According to the vector application method, animals were divided into 3 groups: group 0.35 mL, containing saline solution AAV2/9-LacZ (2 ×1011 vector genome) was injected directly into the myocardium (apex) immediately after reperfusion. Group 0.3 mL contained a cardioplegic solution AAV2/9-LacZ vectors (3 ×1012 vector genome), which was rapidly injected into the aortic root, with the pulmonary trunk clamped. Before transplant the transfected heart was incubated for 25 minutes in iced cardioplegia. A reperfusion system was applied in group 5 mL. For 25 minutes, a cold solution of cardioplegia and AAV2/9-LacZ vectors (5 ×1012 vector genome) was recirculated through the donor heart. Transplanted grafts were explanted after 3 weeks. To detect and to measure marker gene expression, X-gal staining was performed. Results: In groups 0.35 mL and 0.3 mL, higher transfection efficiency was observed compared to group 5 mL (P < .05). While positive-stained myocardia were detected around the injection site in group 0.35 mL, the expression pattern was much more homogenous in group 0.3 mL. Conclusions: Results demonstrate that intracoronary injection of the vectors with the pulmonary trunk clamped leads to the highest and most homogenous distribution of transgene expression in the graft.

Original languageEnglish
Pages (from-to)184-187
Number of pages4
JournalExperimental and Clinical Transplantation
Volume7
Issue number3
StatePublished - Sep 2009
Externally publishedYes

Keywords

  • Cardiac transplant
  • Introcoronary infusion
  • Myocardium
  • Perfusion
  • Transfection

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