A subscription to JoVE is required to view this content. Sign in or start your free trial.
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.
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.
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....
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
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 .......
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.......
None.
....Name | Company | Catalog Number | Comments |
12 Gauge angio-catheter | BD | 382277 | |
14 Gauge angio-catheter | B. Braun | 4251717-02 | |
16 Gauge angio-catheter | B. Braun | 4252586-02 | |
18 Gauge angio-catheter | B. Braun | 4251679-02 | |
20 Gauge angio-catheter | B. Braun | 4252527-02 | |
4-0 silk suture | Surgical Specialties Corp. | SP116 | |
6-0 nylon suture | AD Surgical | S-N618R13 | |
7-0 nylon suture | AD Surgical | S-N718SP13 | |
8-0 nylon suture | AD Surgical | XXS-N807T6 | |
Betadine Spray | Avrio Health L.P | UPC 367618160039 | |
Clippers | VWR | MSPP-023326 | |
Castroviejo micro dissecting spring scissors | Roboz Surgical Instrument Co | RS-5668 | |
Dumont #5 - Fine Forceps | Fine Science Tools | 11254-20 | |
Electrocautery | Macan | MV-7A | |
Endotracheal intubation kit | Kent Scientific | ETI-MSE | |
Forceps | Fine Science Tools | 11027-12 | |
Halsted-mosquito hemostat | Roboz Surgical Instrument Co | RS-7112 | |
Heparin | Fresnius Medical Care | C504701 | |
Insulin syringe | Life Technologies | B328446 | |
Isoflurane | Piramal Critical Care | NDC 66794-017-25 | |
Isopropyl Alcohol Swabs | BD | 326895 | |
Ketamine | Hikma Pharmaceuticals PLC | NDC 0413-9505-10 | |
Dieffenbach Bulldog Clamp | World Precision Instruments | WPI14117 | |
Needle holder/Forceps, Curved | Micrins | MI1542 | |
Needle holder/Forceps, Straight | Micrins | MI1540 | |
Perfadex Plus (Organ Preservation Solution) | XVIVO Perfusion AB | REF# 19950 | |
PhysioSuite | Kent Scientific | PS-MSTAT-RT | Used to check SpO2 and heartbeat |
Retractor | Roboz Surgical Instrument Co | RS-6560 | |
Saline | PP Pharmaceuticals LLC | NDC 63323-186-10 | |
Scissors | Fine Science Tools | 14090-11 | |
SomnoSuite Small Animal Anesthesia System | Kent Scientific | SS-MVG-Module | |
Sterile Cotton Gauze Pad | Fisherbrand | 22-415-469 | |
Surgical Microscope | Leica | M500-N w/ OHS | |
Syringe 5mL | BD | 309646 | |
Vannas-Tubingen Spring Scissors | Fine Science Tools | 15008-08 | |
Xylazine | Korn Pharmaceuticals Corp | NDC 59399-110-20 | |
Yasargil Clamp | Aesculap, Inc | FT351T | Used to clamp bronchus |
Yasargil Clamp | Aesculap, Inc | FT261T | Used to clamp hilum |
Yasargil Clamp Applicator | Aesculap, Inc | FT484T |
This article has been published
Video Coming Soon
ABOUT JoVE
Copyright © 2024 MyJoVE Corporation. All rights reserved