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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

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.

Abstract

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.

Introduction

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 ....

Protocol

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.......

Representative Results

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 .......

Discussion

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.......

Acknowledgements

This research is funded on behalf of the University Hospital Foundation.

....

Materials

NameCompanyCatalog NumberComments
ABL 800 FLEX Blood Gas AnalyzerRadiometer989-963
Adult-Pediatric Electrostatic Filter HME - SmallCovidien352/5877
Allison Lung RetractorPilling341679
Arterial FilterSORIN GROUP01706/03
Backhaus Towel ClampPilling454300
Bovine Serum AlbuminMP biomedicals218057791
Biomedicus PumpMaquetBPX-80
Bronchoscope
Cable Ties – White 12”HUASU InternationalHS4830001
Calcium ChlorideFisher ScientificC69-500G
Cooley Sternal RetractorPilling341162
CUSHING Gutschdressing ForcepsPilling466200
Debakey-Metzenbaum DissectingPilling342202
ScissorsPilling342202
DeBakey Peripheral Vascular ClampPilling353535
Debakey Straight Vascular Tissue ForcepsPilling351808
D-glucoseSigma-AldrichG5767-500G
Drop sucker
Endotracheal Tube 9.0mm CUFDMallinckrodt9590E
Flow TransducerBIO-PROBETX 40
Infusion PumpBaxterAS50
Inspire 7 M Hollow Fiber Membrane OxygenatorSORIN GROUPK190690
Intercept Tubing Connector 3/8" x 1/2"Medtronic6013
Intercept Tubing 1/4" x 1/16" x 8'Medtronic3108
Intercept Tubing 3/8" x 3/32" x 6'Medtronic3506
LaryngoscopeN/AN/ACustom-made with 10-inch blade
Metzenbaum Dissecting ScissorsPilling460420
Medical Carbon Dioxide TankPraxair5823115
Medical Oxygen TankPraxair2014408
Medical Nitrogen TankPraxairNI M-K
Mosquito ClampPilling181816
Harken Auricle Clamp
Organ ChamberTevosol
PlasmaLyte ABaxterTB2544
Poole Suction TubePilling162212
Potassium PhosphateFischer ScientificP285-500G
PERFADEX PlusXVIVO19811
Satinsky ClampPilling354002
ScaleTANITAKD4063611
Silicon Support MembraneTevosol
Sodium BicarbonateSigma-Aldrich792519-1KG
Sodium Chloride 0.9%BaxterJB1324
Sorin XTRA Cell SaverSORIN GROUP75221
Sternal SawStryker6207
Surgical Electrocautery DeviceKls MartinME411
TruWave Pressure TransducerEdwardsVSYPX272
Two-Lumen Central Venous Catheter 7fr X2Arrowg+ardCS-12702-E
Vorse Tubing ClampPilling351377
Willauer-Deaver RetractorPilling341720
Yankauer Suction TubePilling162300
0 ETHIBOND Green 1X36" Endo Loop 0ETHICOND8573
0 PDS II CP-1 2x27”ETHICONZ467H
1 VICRYL MO-4 1x18”ETHICONJ702D
2-0 SILK Black 12" x 18" StrandsETHICONSA77G
4-0 PROLENE Blue TF 1x24”ETHICON8204H
6-0 PROLENE Blue BV 2x30”ETHICONM8776
21-Gauge Needle

References

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Left Lung Orthotopic TransplantationJuvenile Porcine ModelESLPSurgical ManagementEx situ Lung PerfusionIn Vivo ReperfusionThoracotomyPulmonary ArteryPulmonary VeinsBronchusHemiazygos VeinHeparinClampsPigAnesthesiaDissection

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