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Ann Thorac Surg 1995;60:211-212
© 1995 The Society of Thoracic Surgeons

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How To Do It

Revised Technique of Isolated Lung Perfusion in the Rat

Thoracic Oncology Laboratory, Memorial Sloan-Kettering Cancer Center, New York, New York

Accepted for publication April 5, 1995.

	Abstract

Top Footnotes Abstract Introduction Technique Results Comment References

 
Isolated lung perfusion in the rat is a useful research tool. Pulmonary arteriotomy and venotomy repair is required for animal survival. Rat pulmonary vessels are fragile, thin, and small in caliber, making their repair the focal technical point. This technique has been improved. The arteriotomy now is closed with a single suture and the venotomy no longer requires repair. The lung is returned to its original anatomic position then compressed to stop bleeding. These improvements have improved animal morbidity and mortality markedly while reducing operating time.

	Introduction

Top Footnotes Abstract Introduction Technique Results Comment References

 
The technique of isolated rat lung perfusion in vivo was developed in our laboratory to investigate the utility of high-dose regional chemotherapy for the treatment of isolated pulmonary metastases [1]. Isolated lung perfusion in the rat has been technically challenging, requiring endotracheal intubation followed by pulmonary arteriotomy and venotomy for catheterization, drug perfusion, and finally vessel repair. Because the pulmonary vessels are very fragile and measure only 1 mm in diameter in adult rats, technical errors in vessel repair can lead to bleeding, stenosis, occlusion, and ultimately animal death. Significant technical advances have been made in the repair of the pulmonary vessels, which have increased efficiency while decreasing animal morbidity and mortality.

	Technique

Top Footnotes Abstract Introduction Technique Results Comment References

 
Animals used for experiments were treated in compliance with the Animal Welfare Act and ``Guide for the Care and Use of Laboratory Animals'' published by the National Institutes of Health (NIH publication 85-23, revised 1985). Animals were anesthetized with pentobarbital (50 mg/kg) intraperitoneally. Under direct visualization the animals were intubated with a 16-gauge intravenous catheter placed over a guidewire and then placed on a volume ventilator. Ventilation was maintained at a tidal volume of 10 mL/kg with 100% O₂ at a rate of 75 strokes/min. The left side of the chest was shaved, prepared with a 10% povidone-iodine solution, and entered through the fourth intercostal space.

The lung was retracted from the thoracic cavity with a cotton-tipped applicator. The left pulmonary vessels were dissected free. The pulmonary artery and vein were clamped proximally and distally, respectively, with microclips. Under an operating microscope a small incision was created with microscissors in the pulmonary artery and vein. A PE-10 catheter (0.28 mm inner diameter, 0.61 mm outer diameter; Becton Dickinson & Co, Franklin Lakes, NJ) was inserted into the pulmonary artery for infusion. A pulmonary venotomy was performed and the effluent collected by a suction catheter placed in proximity to the venotomy. At the completion of the perfusion, the catheters were removed, the pulmonary artery was repaired with a single transverse suture (9-0 nylon) placed in the center of the arteriotomy, and no venotomy repair was performed (Fig 1).

View larger version (31K): [in this window] [in a new window]   Fig 1. . The lung was retracted from the thoracic cavity with a cotton-tipped applicator. The pulmonary artery and vein were clamped proximally and distally, respectively, with microclips. The artery and vein catheters were removed after lung perfusion was finished. The pulmonary artery was repaired with a single transverse suture (9-0 nylon); the vein was not repaired.

	Results

Top Footnotes Abstract Introduction Technique Results Comment References

 
One hundred eighteen Fischer 344 rats (175 to 250 g) have undergone the revised perfusion, whereas 42 rats were treated with the original method. Four rats (2 in each group) died of an overdose of anesthesia. The complications and mortality after the two repair techniques are compared in Table 1. With the revised technique, there were no perioperative deaths related to uncontrolled bleeding and no significant pulmonary artery stenosis was evident on visual inspection. Blood loss, recorded as chest tube drainage, was consistently less than 1 mL. The time required for pulmonary artery repair and bleeding control was less than 3 minutes. A lung perfusion can now be performed routinely in less than 30 minutes, including 10 minutes of drug perfusion and 5 minutes of saline washout. This enables us to complete 12 to 15 rat lung perfusions safely per day.

	Comment

Top Footnotes Abstract Introduction Technique Results Comment References

Previously, the pulmonary arteriotomy and venotomy were repaired with two or three interrupted 9-0 nylon sutures, respectively. This led to increased tension on the vessels because of vessel shortening. This tension predisposed the vessel to bleeding after the pulmonary circulation was restored. If the bleeding was not controlled, the left pulmonary circulation was reclamped and the vessels were repaired with additional sutures. Tearing of the vessels during repair, especially the pulmonary vein, was not uncommon, leading to uncontrolled bleeding. Placement of additional sutures resulted in vessel stenosis or occlusion, which significantly affects experimental results [3]. Previously, the time required to control bleeding and repair the vessels was extensive and allowed for only four to six lung perfusions in one day. A project would require multiple days of tumor implantation with multiple days of lung perfusions.

Improvements in vessel repair have been made to decrease the technical difficulty of isolated lung perfusion. These include (1) repair of the pulmonary artery with a single suture, (2) no repair of the pulmonary vein, and (3) placement of the perfused lung into the proper anatomic position with pressure placed on the lung, not directly on the vessels. These procedures have decreased the possibility of vessel stenosis and occlusion and eliminated the vessel tension and made it easy to control bleeding. Now, 12 to 15 lung perfusions can be performed in 1 day. A project consisting of 30 rat perfusions now requires only 1 day of tumor implantation with 2 days of lung perfusions.

This improved method of isolated lung perfusion has decreased operative time, morbidity, and mortality. These improvements hopefully will promote rat lung perfusion as an economic, convenient, and useful research tool with more widespread applications.

	Footnotes

Top Footnotes Abstract Introduction Technique Results Comment References

 
Address reprint requests to Dr Burt, Department of Surgery, Memorial Sloan-Kettering Cancer Center, 1275 York Ave, NY 10021.

	References

Top Footnotes Abstract Introduction Technique Results Comment References

 

1. Weksler B, Schneider A, Ng B, Burt M. Isolated single lung perfusion in the rat: an experimental model. J Appl Physiol 1993;74:2736–9.[Abstract/Free Full Text]
2. Minchin RF, Johnston MR, Aiken MA, Boyd MR. Pharmacokinetics of doxorubicin in isolated lungs of dogs and humans perfused in vivo. J Pharmacol Exp Ther 1984;229:193–8.[Abstract/Free Full Text]
3. Wang HY, Ng B, Ahrens C, Burt M. Unilateral pulmonary artery occlusion inhibits growth of pulmonary metastatic sarcoma in the rat. J Surg Oncol 1994;57:183–6.[Medline]

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This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Personal Folders Download to citation manager Author home page(s): Jeffrey L. Port Michael E. Burt Permission Requests Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wang, H.-Y. Articles by Burt, M. E. Search for Related Content PubMed PubMed Citation Articles by Wang, H.-Y. Articles by Burt, M. E.