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This protocol describes a juvenile porcine model of orthotopic left lung allotransplantation designed for use with ESLP research. Focus is made on anesthetic and surgical techniques, as well as critical steps and troubleshooting.
Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. However, lung transplantation is limited by a shortage of available donor organs. As such, there is high waitlist mortality. Ex situ lung perfusion (ESLP) has increased donor lung utilization rates in some centers by 15%-20%. ESLP has been applied as a method to assess and recondition marginal donor lungs and has demonstrated acceptable short- and long-term outcomes following transplantation of extended criteria donor (ECD) lungs. Large animal (in vivo) transplantation models are required to validate ongoing in vitro research findings. Anatomic and physiologic differences between humans and pigs pose significant technical and anesthetic challenges. An easily reproducible transplant model would permit the in vivo validation of current ESLP strategies and the preclinical evaluation of various interventions designed to improve donor lung function. This protocol describes a porcine model of orthotopic left lung allotransplantation. This includes anesthetic and surgical techniques, a customized surgical checklist, troubleshooting, modifications, and the benefits and limitations of the approach.
Lung transplantation is the preeminent long-term treatment for end-stage lung disease. Over 4,600 lung transplantations are performed worldwide annually1. However, lung transplantation currently has significant limitations. For one, the necessity for organs continues to eclipse available donors. Despite rates of lung transplantation increasing every year since 2012 due to the combined effects of more candidates being listed for transplant, an increase in the number of donors, and improved use of recovered organs, the transplant waitlist mortality has not decreased significantly2. Organ quality concerns represent another ....
All the procedures were performed in compliance with the guidelines of the Canadian Council on Animal Care and the guide for the care and use of laboratory animals. The protocols were approved by the institutional animal care committee of the University of Alberta. This protocol has been applied in female juvenile Yorkshire pigs between 35-50 kg. Pigs are pathogen-free, food-grade specimens. They are purchased from the Swine Research and Technology Centre in Edmonton, AB, Canada (https://srtc.ualberta.ca). All individual.......
All of the results are in the context of 4 h of reperfusion following 12 h of NPV-ESLP16. During lung explant, there are several clinical outcomes to anticipate (Figure 3). Typically, the pig will remain hemodynamically stable following a successful left lung explantation but may require a low dose infusion of phenylephrine (dose range: 2-10 mg/h) due to a vasodilatory response to surgery. Heart rate should target approximately 100-120 bpm, respiratory rate (RR) 8-30 .......
Several critical surgical steps are involved in this protocol, and troubleshooting is needed to ensure successful transplantation and lung assessment. Juvenile porcine lungs are incredibly delicate compared to adult human lungs, so the operating surgeon must be cautious when handling porcine lungs. This is especially true after a 12 h run of ESLP as the organ will have taken on fluid volume and be susceptible to injury from excessive manipulation. Any undue pressure will cause atelectasis or trauma to the experimental lu.......
This research is funded on behalf of the University Hospital Foundation.
....Name | Company | Catalog Number | Comments |
ABL 800 FLEX Blood Gas Analyzer | Radiometer | 989-963 | |
Adult-Pediatric Electrostatic Filter HME - Small | Covidien | 352/5877 | |
Allison Lung Retractor | Pilling | 341679 | |
Arterial Filter | SORIN GROUP | 01706/03 | |
Backhaus Towel Clamp | Pilling | 454300 | |
Bovine Serum Albumin | MP biomedicals | 218057791 | |
Biomedicus Pump | Maquet | BPX-80 | |
Bronchoscope | |||
Cable Ties – White 12” | HUASU International | HS4830001 | |
Calcium Chloride | Fisher Scientific | C69-500G | |
Cooley Sternal Retractor | Pilling | 341162 | |
CUSHING Gutschdressing Forceps | Pilling | 466200 | |
Debakey-Metzenbaum Dissecting | Pilling | 342202 | |
Scissors | Pilling | 342202 | |
DeBakey Peripheral Vascular Clamp | Pilling | 353535 | |
Debakey Straight Vascular Tissue Forceps | Pilling | 351808 | |
D-glucose | Sigma-Aldrich | G5767-500G | |
Drop sucker | |||
Endotracheal Tube 9.0mm CUFD | Mallinckrodt | 9590E | |
Flow Transducer | BIO-PROBE | TX 40 | |
Infusion Pump | Baxter | AS50 | |
Inspire 7 M Hollow Fiber Membrane Oxygenator | SORIN GROUP | K190690 | |
Intercept Tubing Connector 3/8" x 1/2" | Medtronic | 6013 | |
Intercept Tubing 1/4" x 1/16" x 8' | Medtronic | 3108 | |
Intercept Tubing 3/8" x 3/32" x 6' | Medtronic | 3506 | |
Laryngoscope | N/A | N/A | Custom-made with 10-inch blade |
Metzenbaum Dissecting Scissors | Pilling | 460420 | |
Medical Carbon Dioxide Tank | Praxair | 5823115 | |
Medical Oxygen Tank | Praxair | 2014408 | |
Medical Nitrogen Tank | Praxair | NI M-K | |
Mosquito Clamp | Pilling | 181816 | |
Harken Auricle Clamp | |||
Organ Chamber | Tevosol | ||
PlasmaLyte A | Baxter | TB2544 | |
Poole Suction Tube | Pilling | 162212 | |
Potassium Phosphate | Fischer Scientific | P285-500G | |
PERFADEX Plus | XVIVO | 19811 | |
Satinsky Clamp | Pilling | 354002 | |
Scale | TANITA | KD4063611 | |
Silicon Support Membrane | Tevosol | ||
Sodium Bicarbonate | Sigma-Aldrich | 792519-1KG | |
Sodium Chloride 0.9% | Baxter | JB1324 | |
Sorin XTRA Cell Saver | SORIN GROUP | 75221 | |
Sternal Saw | Stryker | 6207 | |
Surgical Electrocautery Device | Kls Martin | ME411 | |
TruWave Pressure Transducer | Edwards | VSYPX272 | |
Two-Lumen Central Venous Catheter 7fr X2 | Arrowg+ard | CS-12702-E | |
Vorse Tubing Clamp | Pilling | 351377 | |
Willauer-Deaver Retractor | Pilling | 341720 | |
Yankauer Suction Tube | Pilling | 162300 | |
0 ETHIBOND Green 1X36" Endo Loop 0 | ETHICON | D8573 | |
0 PDS II CP-1 2x27” | ETHICON | Z467H | |
1 VICRYL MO-4 1x18” | ETHICON | J702D | |
2-0 SILK Black 12" x 18" Strands | ETHICON | SA77G | |
4-0 PROLENE Blue TF 1x24” | ETHICON | 8204H | |
6-0 PROLENE Blue BV 2x30” | ETHICON | M8776 | |
21-Gauge Needle |
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