In situ gene transfer into rat auxiliary liver transplant

Transplantation. 1997 Dec 15;64(11):1537-41. doi: 10.1097/00007890-199712150-00006.

Abstract

Background: A replication-defective retrovirus BAG vector was tested for in situ delivery of the beta-galactosidase gene to auxiliary liver transplant in a rat model.

Methods: The BAG vector, which was shown to be effective in genetic transduction of cultured NIH/3T3 cells, was produced in a psi2 packaging cell and later amplified in a selected PA317 clone. Hepatocyte replication was induced by one-third hepatectomy of the donor liver, and the procedure was followed by auxiliary partial liver transplantation. Twenty-four hours after hepatic induction or transplantation, viral supernatant at 37 degrees C was perfused into the liver graft via the portal vein during a temporary occlusion of the graft portal vein.

Results: All animals survived the transplantation procedures and were killed at specified time intervals. Histochemical staining of the liver graft specimens indicated the expression of beta-galactosidase in the gene transferred group but not in the control animals. As demonstrated by polymerase chain reaction assay, the proviral beta-galactosidase sequence was present in the graft specimens, but absent from all other tissues tested.

Conclusions: In short, the retrovirus BAG vector can be useful for in situ delivery of foreign genes to liver graft in transplantation and other clinical settings, providing a simple, consistent, and reliable alternative in hepatic gene therapy experiments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Cell Line
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Vectors
  • Liver Transplantation / methods*
  • Male
  • Mice
  • Polymerase Chain Reaction
  • Rats
  • Rats, Inbred Lew
  • Retroviridae
  • Transfection
  • beta-Galactosidase / genetics*

Substances

  • beta-Galactosidase