title: Ex vivo hypothermic recirculatory adenoviral gene transfer to the transplanted pig heart.
abstract: BACKGROUND
  To facilitate the application of adenoviral gene therapy in clinical heart transplantation, we developed an ex vivo hypothermic recirculatory adenoviral gene transfer method to the transplanted pig heart.
  
  
  METHODS
  Experimental animals were assigned into three groups; controls, 1x10(8) plaque-forming units (pfu)/ml group and 1x10(9) pfu/ml group. During the 30 min gene transfer perfusion, 200 ml of University of Wisconsin solution containing the adenoviral vector was recirculated through the coronary vessels. The myocardial temperature was maintained below 4 degrees C and the perfusion pressure was adjusted at 50 mmHg.
  
  
  RESULTS
  Cardiac myocyte transduction efficiencies in the 1x10(8) pfu/ml group were 0.04% and 0.07%, whereas transduction efficiencies in the 1x10(9) pfu/ml group were widely distributed from 0.45% to 22.62%. The gene transduction efficiency increased with the virus titer. Additionally, no difference in the transduction efficiency was observed between different segments of the left ventricle. The current gene transfer method at 1x10(9) pfu/ml of adenovirus titer enabled homogeneous gene transduction into the transplanted pig heart up to a maximum of 22.62%.
  
  
  CONCLUSIONS
  This model can be applied to a large isolated heart and will greatly facilitate the investigation of gene therapy in large animal models of heart transplantation.
authors.affiliation: Mayo Clinic William J. von Liebig Transplant Center, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Keiji
authors.initials: K
authors.lastname: Oi
authors.forename: William R
authors.initials: WR
authors.lastname: Davies
authors.forename: Henry D
authors.initials: HD
authors.lastname: Tazelaar
authors.forename: Kent R
authors.initials: KR
authors.lastname: Bailey
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Federspiel
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Russell
authors.forename: Christopher G A
authors.initials: CG
authors.lastname: McGregor
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D003080:Cold Temperature; D004487:Edema / Q000209:etiology; D018014:Gene Transfer Techniques / Q000295:instrumentation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016027:Heart Transplantation / Q000009:adverse effects / Q000379:methods / Q000473:pathology; D066298:In Vitro Techniques; D010477:Perfusion; D034421:Sus scrofa; D014161:Transduction, Genetic; D014184:Transplantation, Homologous