| Surgical organ perfusion method for somatic gene transfer: An experimental study on gene transfer into the kidney, spleen, lung and mammary gland | ||
|---|---|---|
| Prev | Next | |
A total of 61 experimental operations were made on the animals, i.e. rabbits, pigs and goats. A summary of the vector delivery methods and the overall results is presented in Table 3.
Table 3. Summary of animal experiments used in this study.
| Animal | n | Organ | Method for vector delivery | Result |
|---|---|---|---|---|
| Rabbit | 4 | Kidney | Intra-arterial infusion | No expression |
| Pig | 11 | Kidney | Intra-arterial infusion with vasodilative agents | No expression |
| Pig | 2 | Spleen | Intra-arterial infusion | No expression |
| Pig | 2 | Kidney | Ex vivo perfusion at room temperature | No expression |
| Pig | 4 | Kidney | Ex vivo perfusion at body temperature | Expression up to 80% of glomeruli |
| Pig | 1 | Lung | Ex vivo perfusion at body temperature | Expression in vascular and non-vascular cells |
| Pig | 13 | Kidney | In vivo perfusion | Expression up to 75% of glomeruli |
| Pig | 10 | Spleen | In vivo perfusion | Expression in the perifollicular area |
| Pig | 11 | Lung | In vivo perfusion | Expression mostly in the airway and alveolar epithelial cells |
| Goat | 3 | Mammary gland | In vivo perfusion | hGh secretion in the goats milk |
| Total | 61 |
The first set of experiments involving injection of the virus into the renal arteries of four rabbits and eleven farm pigs in vivo did not lead to any successful gene transfer into renal cells, as it appeared four days after the injection. β -galactosidase gene expression was only seen in a few scattered cells in the renal cortex, while no signs of expression were seen in the glomeruli (data not shown). The adjuvant vasodilative agents used in order to diminish possible intrarenal vascular resistance immediately prior to the injection of the virus had no visible effect on the efficiency of gene transfer (Table 4).
Table 4. Experimental intra-arterial infusions of virusparticles into renal artery in conjunction with various vasodilative agents.
| Experimental animal | Vasodilative agent | Number of virusparticles (pfu ml–1) |
|---|---|---|
| A1 | None | Ad 2x109 |
| A2 | Papaverin 30 mg | Ad 4x109 |
| A3 | Papaverin 20 mg | Ad 4x109 |
| A4 | Papaverin 10 mg | Ad 4x109 |
| A5 | Alprostadil 5 µg | Ad 4x109 |
| A6 | Verapamil 2 mg | Ad 4x109 |
| A7 | Enalapril 0.5 mg | Ad 4x109 |
| A8 | Lidocain 100 mg | Ad 4x109 |
| A9 | Papaverin 60 mg | Ad 4x109 |
| A10 | NaCl 10 ml | Ad 4x109 |
| A11 | Papaverin 20 mg + NaCl 10 ml | Ad 4x109 |
| pfu= plaque forming unit | ||
Infusion of the adenoviral solution into the splenic artery did not result in any notable β -galactosidase gene expression at four days after the intra-arterial infusions in two experiments.