Design and Implementation of a Rat Ex Vivo Lung Perfusion Model

Summary

Ex vivo lungs are useful for a variety of experiments to collect physiological data while excluding the confounding variables of in vivo experiments. Commercial setups are often expensive and limited in the types of data they can collect. We describe a method for building a fully modular setup, adaptable for various study designs.

Abstract

Ex vivo lung preparations are a useful model that can be translated to many different fields of research, complementing corresponding in vivo and in vitro models. Laboratories wishing to use isolated lungs need to be aware of important steps and inherent challenges to establish a setup that is affordable, reliable, and that can be easily adapted to fit the topic of interest. This paper describes a DIY (do it yourself) model for ex vivo rat lung ventilation and perfusion to study drug and gas effects on pulmonary vascular tone, independent of changes in cardiac output. Creating this model includes a) the design and construction of the apparatus, and b) the lung isolation procedure. This model results in a setup that is more cost-effective than commercial alternatives and yet modular enough to adapt to changes in specific research questions. Various obstacles had to be resolved to ensure a consistent model that is capable of being used for a variety of different research topics. Once established, this model has proven to be highly adaptable to different questions and can easily be altered for different fields of study.

Introduction

Ex vivo lung perfusion (EVLP) techniques¹ have seen a rise in usage in the past decade as a means of studying lung transplantations², ischemia/reperfusion³, lung metabolism⁴, and immune responses⁵. Isolated, but intact, ventilated and perfused lungs offer the critically important ability to directly assess the response of the lungs, including the pulmonary vasculature, to potential interventions and/or therapeutics without potential confounders, such as neuronal and hormonal input or changing hemodynamics in vivo. At the same time, ....

Protocol

The in vivo portion of the experiments (from general anesthesia to euthanasia) requires prior approval by the respective Institutional Animal Care and Use Committee (IACUC). All procedures described herein were approved (protocol number M1700168) by the IACUC at Vanderbilt University Medical Center, Nashville, Tennessee, and were performed in compliance with the ARRIVE guidelines¹⁴. Prior to experimentation, all the rats were housed in the institute's animal care facility, with free a.......

Discussion

More than 100 experiments have been successfully performed in our lab using this setup. The modular design of this customized setup gave great flexibility to potential changes in experimental requirements. While other setups utilize a deoxygenator¹⁸ to mimic constant oxygen consumption and CO₂ production by end organs, this simplified model did not employ this feature, due to the focus on studying the effects of different gas compositions on pulmonary vascular tone. This approach, in wh.......

Materials

Name	Company	Catalog Number	Comments
1,000 mL Glass Beaker	Pyrex, Chicago, IL
1,500 mL Glass Beaker	Pyrex, Chicago, IL
Air Trap Compliance Chamber	Radnoti	130149
Bioamplifiers	CWE Inc	BPM-832
Clamps	Fisher Scientific	S02626
DAQ (Data Acquisition)	National Instruments, Austin, TX	NI USB-6343
Gas Mixer	CWE Inc, Ardmore, PA	GSM-4
Heating Coil	Radnoti, Covina, CA	158822
Heating Plate	Thermo Fisher Scientific, Waltham, MA	11-100-49SH
Heparin	Pfizer	W63422
LabVIEW Full Development System 2014	National Instruments
Pentobarbital	Diamondback Drugs	G2270-0235-50
pH700 Probe	OAKTON, Vernon Hills, IL	EW-35419-10
Polystat Water Bath	Cole-Parmer	EW-12121-02
Rodent Ventilator	Harvard Apparatus, Holliston, MA	Model 683
Roller Pump	Cole-Parmer, Wertheim, Germany	Ismatec REGLO Digital MS 2/8
Sprague Dawley Rat	Charles River, Wilmington, MA	Strain code 001
VetScan i-STAT	Abraxis, Chicago, IL	i-STAT 1