Sign In

A subscription to JoVE is required to view this content. Sign in or start your free trial.

In This Article

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

Summary

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

Abstract

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Introduction

For over three decades, researchers have been modifying and improving rat lung transplantation models so that the data generated are more consistent and more reflective of the actual clinical condition. In our laboratory's time performing this model, we have determined four areas of improvement: cuffing techniques for anastomoses, identification of the recipient's 4th intercostal space, lung inflation and wound closure during the recipient's procedure, and the harvesting of samples for analysis.

Cuffing technique modifications for anastomoses can improve the entire transplantation procedure by shortening handling time....

Protocol

Male Sprague-Dawley rats (180-270 g body weight) were purchased commercially (e.g., Envigo) and were housed under pathogen-free conditions at The Ohio State University Animal Facility. All procedures were humanely performed according to the NIH and the National Research Council's Guide for the Humane Care and Use of Laboratory Animals and with the approval of The Ohio State University Institutional Animal Care and Use Committee (IACUC Protocol # 2012A00000135-R2).

1. Initial setup

    .......

Representative Results

In order to measure pulmonary edema, the wet-to-dry weight ratio was calculated. The donor's native lobe, the transplanted lobe, and the recipient's native lobe were collected as described in the protocol and weighed immediately for wet weight, dried at 60 °C for 48 h, and then weighed again for the dry weight. An increased wet-to-dry weight ratio would be indicative of pulmonary edema. Our results indicate that the transplanted lobe did have a significant increase in wet-to-dry .......

Discussion

In this report, we have intervened at several critical steps in a rat lung transplantation protocol to optimize the procedure. While various cuffing techniques for rat lung transplantation has been reported1,2,3,4,5,6,7,8,9.......

Acknowledgements

None.

....

Materials

NameCompanyCatalog NumberComments
12 Gauge angio-catheterBD382277
14 Gauge angio-catheterB. Braun4251717-02
16 Gauge angio-catheterB. Braun4252586-02
18 Gauge angio-catheterB. Braun4251679-02
20 Gauge angio-catheterB. Braun4252527-02
4-0 silk sutureSurgical Specialties Corp.SP116
6-0 nylon sutureAD SurgicalS-N618R13
7-0 nylon sutureAD SurgicalS-N718SP13
8-0 nylon sutureAD SurgicalXXS-N807T6
Betadine SprayAvrio Health L.PUPC 367618160039
ClippersVWRMSPP-023326
Castroviejo micro dissecting spring scissorsRoboz Surgical Instrument CoRS-5668
Dumont #5 - Fine ForcepsFine Science Tools11254-20
ElectrocauteryMacanMV-7A
Endotracheal intubation kitKent ScientificETI-MSE
ForcepsFine Science Tools11027-12
Halsted-mosquito hemostatRoboz Surgical Instrument CoRS-7112
HeparinFresnius Medical CareC504701
Insulin syringeLife TechnologiesB328446
IsofluranePiramal Critical CareNDC 66794-017-25
Isopropyl Alcohol SwabsBD326895
KetamineHikma Pharmaceuticals PLCNDC 0413-9505-10
Dieffenbach Bulldog ClampWorld Precision InstrumentsWPI14117
Needle holder/Forceps, CurvedMicrinsMI1542
Needle holder/Forceps, StraightMicrinsMI1540
Perfadex Plus (Organ Preservation Solution)XVIVO Perfusion ABREF# 19950
PhysioSuiteKent ScientificPS-MSTAT-RTUsed to check SpO2 and heartbeat
RetractorRoboz Surgical Instrument CoRS-6560
SalinePP Pharmaceuticals LLCNDC 63323-186-10
ScissorsFine Science Tools14090-11
SomnoSuite Small Animal Anesthesia SystemKent ScientificSS-MVG-Module
Sterile  Cotton Gauze PadFisherbrand22-415-469
Surgical MicroscopeLeicaM500-N w/ OHS
Syringe 5mLBD309646
Vannas-Tubingen Spring ScissorsFine Science Tools15008-08
XylazineKorn Pharmaceuticals CorpNDC 59399-110-20
Yasargil ClampAesculap, IncFT351TUsed to clamp bronchus
Yasargil ClampAesculap, IncFT261TUsed to clamp hilum
Yasargil Clamp ApplicatorAesculap, IncFT484T

References

  1. Mizuta, T., Kawaguchi, A., Nakahara, K., Kawashima, Y. Simplified rat lung transplantation using a cuff technique. Journal of Thoracic and Cardiovascular Surgery. 97 (4), 578-581 (1989).
  2. Zhai, W., et al. Simplified rat lung....

Explore More Articles

Rat Lung TransplantationWarm Ischemia reperfusion InjurySurgical TechniqueCuffing MethodAngio catheterThoracic IncisionEndotracheal IntubationHeparin AdministrationMolecular MechanismsTherapeutic Interventions

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved