A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Summary

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

Abstract

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies' findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Introduction

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure¹. Although ischemic times of up to four hours are tolerated adequately, an ischemic time of greater than six hours is associated with inferior outcomes². Primary graft dysfunction remains the principal cause of early morbidity and mortality following transplantation^2,3. The causes of primary graft dysfunction are multifactorial and include the use of marginal organs, recipient pulmonary vascular disease, ....

Protocol

The institutional animal care committee approved all experimental protocols and animals were treated following the “Guide for the Care and Use of Laboratory Animals” prepared by the Institute of Laboratory Animal Resources, National Research Council, 1996. Male Yorkshire pigs (40–50 kg) were used to perform the orthotopic heart transplants (animal size can vary according to the investigators discretion and experimental goals).

1. Donor Procedure

1. Anesthetic induction and animal preparation:
   1. Premedicate the animal using an intramuscular injection of Ketamine (20 mg/kg), Midazolam (0.3 mg/kg), and Atro....

Results

This pre-clinical model has been used successfully since 1994^5,6,8,9. Table 1 demonstrates representative results from pressure-volume relationships and echocardiographic parameters taken at baseline, and 3 h post-transplantation in a set of 5 experiments. Although we see a decrease in myocadial contractility following transplantation, this was not statistically significant.

Discussion

This manuscript describes a large-animal pre-clinical model of orthotopic heart transplantation. Various small animal models of heterotopic heart transplantation have been successfully used to study the effects of novel treatments to improve organ preservation and decrease ischemia-reperfusion injury^11,12,13. Additionally, small animal models permit survival models and long-term follow-up to investigate the development of reject.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Amiodarone			Purchased from institutional pharmacy
Angiocath 20G	BD	381704
Calcium Chloride 1g/10ml			Purchased from institutional pharmacy
Cardioplegia solution			This should be chosen at the investigators discretion.
Cautery Pencil	Covidien	E2515H
Central Venous Catheter double-lumen	Cook Medical	C-UDLM-501J-LSC
CPB pack	Medtronic		Custom-made cardiopulmonary bypass perfusion circuit.
D5W 5% 250ml	Baxter	JB1064
DLP Aortic Root Cannula/stabber	Medtronic	12218
DLP single-lumen venous cannula (24F or 28F)			This should be chosen at the investigators discretion.
Dobutamine			Purchased from institutional pharmacy
Electrode Polyhesive	Covidien	E7507
EOPA arterial cannula (17F or 21F)			This should be chosen at the investigators discretion.
Epinephrine			Purchased from institutional pharmacy
Eppendorf Tubes, 1.5 mL	Sarstedt	72.690.001
Gloves, nitrile, medium	Fischer	27-058-52
Heparin 1000 IU/ml			Purchased from institutional pharmacy
Ketalean (Ketamine) inj. 100mg/ml, 50ml/vial	Health Canada		Requires health canada approval
Lidocaine/Xylocaine 1%			Purchased from institutional pharmacy
Magnesium Sulfate 5g/10ml			Purchased from institutional pharmacy
Midazolam inj. USP 5mg/ml vial/10ml	Health Canada		Requires Health canada approval
MPS Quest delivery disposable pack	Quest medical	5001102-AS
NACL 0.9% 1L	Baxter	JB1324
Organ Bag	CardioMed	2990
Pipette Tips, 1 mL	Fisherbrand	02-707-405
Propofol 1mg/ml			Purchased from institutional pharmacy
Rocuronium			Purchased from institutional pharmacy
Set Admin Prim NF PB W/Checkvalve	Smith Medical	21-0442-25	Intravenous infusion pump line. Researchers should choose infusion lines compatible with the infusion pump available at their facilities
Set Intro Sheath 8.5FRx 10CM	Arrow	SI-09880
Sofsilk 0 wax coated	Covidien	S316
Solumedrol 500mg/5ml			Purchased from institutional pharmacy
Suction tip	Covidien	8888501023
Suction Tubing 1/4" x 120"	Med-Rx	70-8120
Suture 5.0 Prolene BB	Ethicon	8580H
Suture Prolene Blum 4-0 SH 36	Ethicon	8521H
Sutures 2.0 Prolene Blu M SH	Ethicon	8523H
Sutures BB 4.0 Prolene	Ethicon	8881H
Tracheal Tube, 6.5mm	Mallinckrodt	86449