Successful long-term storage of rat limbs

Abstract

The effect of simple cold storage in Euro-Collins solution on prolonging successful preservation time was examined on an established rat limb transplant model. Limbs amputated at mid-thigh level were simply immersed in the solution and stored in a refrigerator at 4 ° C. After varying periods of cold storage, the limb was orthotopically grafted to another isogeneic rat using microsurgical techniques. The survival rates for 12, 24, 48 and 72 hours' storage were 100%, 90%, 67% and 50% respectively, and the longest time of successful preservation achieved was 78 hours. Survival rates were greatly decreased when amputated limbs were initially perfused with Euro-Collins solution and immersed in the same solution. In a third set of experiments, the limbs were preserved by a conventional method of cold storage in air, and the survival rates were decreased even further. Evaluation by measuring postoperative paw swelling and microscopic examination of the grafts further confirmed the effectiveness of the simple immersion method without perfusion. R6sum6. Nous avons dtudi~, sur la patte de rat, choisie comme modble de transplant, l'effet du simple stockage au froid dans la solution Euro-Collins sur la prolongation du temps de conservation. Les pattes, amputees it mi-cuisse, ont dt~ simplement plong~es dans la solution, puis conservdes dans un rdfrigdrateur it la tempdrature de 4°C. Apr~s des durdes variables de stockage au froid les pattes ont dtd greffdes sur d'autres rats isog~niques, de fafon de survie des transplants aprbs 12, 24, 48 et 72 heures de conservation fut respectivement de 100%, 90%, 67% et 50%, et la plus longue dur(e de conservation efficace fut de 78 heures. Les taux de survie ont diminud de fafon importante lorsque les transplants Ctaient d'abord perfusds avec la solution Euro-Collins puis immergds dans cette m6me solution. Dans un troisi~me groupe d'expdriences, les pattes amputees furent conservdes de fafon traditionnelle par stockage au froid it l'air, et les taux de survie des greffes diminu~rent encore plus. L'dvaluation par la mesure du gonflement de la patte greffde ainsi que l' examen microscopique ont confirmd l'efficacit~ de la simple immersion sans petfusion.

Assessing viability of extracorporeal preserved muscle transplants using external field stimulation: a novel tool to improve methods prolonging bridge-to-transplantation time

“…In the field of reconstructive medicine with replantation of extremities or free tissue transplantation, the use of EP did not have any impact on clinical practice, but has been the focus of transplantational research in the last decades 4 7 8 34 . The superiority of perfusion over ischemic cold storage has already been shown for skeletal muscles 35 .…”

“…In physical and anatomical studies it could be demonstrated that irreversible muscle cell damage starts after three hours of ischemia and is nearly complete after six hours 3 . Attempts to prolong tissue viability and functionality that have been delevoped and thoroughly tested in the last decade are quite diversified, including cold storage, warm storage, perfusion with blood or blood-like substitutes 4 5 6 7 8 9 10 11 12 13 14 . Current standard conservation protocols for organs and amputated extremities are constricted to ischemic cold storage.…”

“…To assess the efficacy of developed conservation protocols, numerous methods have been developed to evaluate tissue viability, functionality or survival probability. Most are located in the field of histology 4 5 8 11 17 and immunohistochemistry 5 9 11 17 18 . The vast majority of these methods demands complex, costly and time-consuming protocols and delivers narrow, localised results from a retrospective point of view.…”

Tissue conservation for transplantation

“…During the process of cryopreservation, cells are protected with the use of several cryoprotective substances. There are many studies in the literature testing different substances/solutions for composite tissue preservation purposes and reporting different results (Supplemental Table 4) [76], [77], [78], [79], [80], [81], [82], [83], [84], [85].…”

Cold preservation of rat osteochondral tissues in two types of solid organ preservation solution, culture medium and saline

“…University of Wisconsin (UW) solution has enabled longer-term preservation of solid organs when compared with EC solution (Southard and Belzer 1995) and could therefore also be suitable for osteochondral preservation, but has also not been tested in this respect. Recently, cold preservation of musculo-skeletal tissues such as whole rat limbs (Yokoyama et al 1996;Arai et al 1993;Suzuki 1991), and canine vertebral body-disc units (Wakabayashi and Matsuzaki 1995) in EC solution or UW solution has been reported. Because UW and EC solutions have both already been employed for preservation of solid organs in clinical practice for many years, these solutions are readily available for clinical use for the preservation of osteochondral allografts.…”