BACKGROUNDOrgan preservation keeps the quality of the organs under prolonged ischemia. Continuous machine perfusions are gaining an important position in clinical research and practice. The aim of this study was to evaluate the protective effect of continuous hypothermic machine perfusion transport system (AirdriveTM) on cold ischemic injury of canine kidney.

METHODSTen kidneys of five healthy preserving canines were taken out after general anesthesia. Five kidneys were stored using common cold preservation (CCP group) by immersing it in the organ preservation solution, mixed with water and ice, and kept in a cold room at 4°C. The other five kidneys were stored using continuous machine perfusion preservation (CMP group) and were placed into the Airdrive(TM) continuous machine perfusion device at room temperature. The renal tissues were examined by histopathology, electron microscopy, and mitochondrial activity check at different time points.

RESULTSHistologic sections showed that the structures of the ten renal tissues were similar during the first 24 hours. After 48 hours, the CCP group showed more pronounced changes, as the renal tubular epithelial cells were more obvious than those in the glomeruli. Oxygen consumption rate of state III and IV respiration in the CCP group decreased after 12-48 hours and increased at 48 hours, respectively, when compared to the CMP group (P < 0.05). Cortex respiratory control ratio and phosphorus oxygen ratio were significantly higher in the CMP group at 48 hours.

CONCLUSIONWith prolonged storage time, the effect of continuous hypothermic machine perfusion transport system is better than that of common cold preservation on canine kidney.

Animals ; Dogs ; Kidney ; Kidney Transplantation ; Male ; Organ Preservation ; methods ; Organ Preservation Solutions

OBJECTIVETo investigate the protective effect of luteolin on isolated rat heart in hypothermic preservation.

METHODSForty male SD rats were randomly divided into 4 groups (n = 10): control group, luteolin low-dose group (7.5 micromol/L), middle-dose group (15 micromol/L) and high dose group (30 micromol/L). Langendorff model of isolated rat heart was used. After 30 min basal perfusion, the hearts were stored in University of Wisconsin solution (UW solution) at 4 degrees C with luteolin (7.5, 15 and 30 micromol/L) or without luteolin for 12 h and followed by 60 min reperfusion. The recovery of cardiac contractile and diastolic function, coronary flow (CF), creatine kinase (CK) leakage in the coronary effluent, myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULTSThe results revealed that luteolin improved the recovery of left ventricular peak systolic pressure and +/- dp/dtmax dose-dependently and increased coronary flow. The leakage of creatine kinase in the coronary effluent was significantly reduced in luteolin-added hearts. Impairment of myocardial ultrastructure after 12 h hypothermic preservation was obviously alleviated in hearts luteolin-added group compared with that in control group. There were no differences between the groups in myocardial water contents.

CONCLUSIONLuteolin as a supplementation in cardiac preservation solution can significantly improve the hypothermic preservation effects on rat heart and have myocardial protection effect, especially in luteolin-added with 30 micromol/L.

Animals ; Cryopreservation ; In Vitro Techniques ; Luteolin ; pharmacology ; Male ; Myocardium ; Organ Preservation ; methods ; Organ Preservation Solutions ; Rats

In this brief review, some key issues related to vitreous cryopreservation of living tissues (natural or engineered), including cells, embryos, tissues, organs, and engineered tissues, are outlined. The principle of vitreous cryopreservation for the biological activity and functionality is demonstrated. The procedures of cooling/ rewarming, composition and function of optimal cryoprotectants, and their effects on bioproducts are described. Vitrification could, therefore, prove to be a useful and effective method of bioproduct cryopreservation for a long period of time, particularly for organized tissues and organs. However, the toxicity of the cryoprotective agents and the devitrification occurring during the rewarming process need additional investigations. Several key areas of research on vitrification are also addressed.
Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Humans ; Organ Preservation ; methods ; Tissue Preservation ; methods

OBJECTIVETo investigate the effective method of preserving composite facial allograft so as to attenuate ischemic injury.

METHODSThe composite facial allografts were harvested from dog, perfused and preserved with 4 degrees C physiological sodium chloride and UW solution respectively. Immediately after the removal of the flap, after 12, 24, 48 h of preservation, MTT assay was used to determine the viability of several kinds of tissue, including skin, mucosa, muscle, bleed vessel, nerve and gland. The results of the two groups were compared in term of viability percentage. The pathology of several tissues were observed after 24 and 48 h of storage.

RESULTSThe viability percentage of every tissue conserved in UW solution for 48 hours was more than 75%. There was significant difference between physiological sodium chloride group and UW group (P < 0.05). Some changes, including Porous arrangement of fibers in connective tissue of skin and mucosa, hyalinization of tissue around the hair follicle and edema of cell in hair follicle, enlargement of space between muscle bundles and unclearness of boundary of acinus could be seen in physiological sodium chloride group while no significant change in UW group.

CONCLUSIONSUW solution could be considered as preservation solution for composite facial allograft.

Adenosine ; Allopurinol ; Animals ; Dogs ; Face ; Female ; Glutathione ; Insulin ; Male ; Organ Preservation Solutions ; Raffinose ; Tissue Preservation ; methods ; Transplantation, Homologous

The quality of a donor liver after cardiac death is closely associated with energy metabolism during preservation. Ex vivo mechanical perfusion has broad application prospects because this technique can help energy metabolism and repair ischemia injury of donors' livers. Some core issues are presented in this review in order to provide references for propelling secure application of liver transplantation based on donation after cardiac death.
Death ; Humans ; Liver ; Liver Transplantation ; Organ Preservation ; methods ; Perfusion ; methods ; Warm Ischemia ; adverse effects

Organ transplantation is an effective approach for the treatment of end-stage organ failures. Currently, the donor organs used for clinical transplantation are all preserved at above-zero temperatures. These preservation methods are well-established and simple but the storage time lasts for only 4-12 h. Some researchers tried to extend the organ storage time by improving protectant and HLA matching to raise the use of stored organs and prolong the long-term survival of organs. These efforts still fall short of the clinical demand for organ transplantation. Moreover, a great many organs were wasted due to limited storage time, HLA mismatch, patients' conditions or distance involved. Therefore, preserving organs for several weeks or even months and establishing Organ Bank are the tough challenges and have become a shared goal of global scholars. This article reviews some issues involved in the cryopreservation of organs, such as use of cryoprotecting agents, freezing and thawing methods in the cryopreservation of hearts, kidneys and other organs.
Cryopreservation ; methods ; Cryoprotective Agents ; adverse effects ; pharmacology ; Humans ; Organ Preservation ; methods

OBJECTIVETo summarize the experience of donor liver procurement and preparation in liver transplantation.

METHODSOne hundred and twenty-six cases of donor liver and kidney procurement and 105 cases of donor liver preparation from August, 2004 to December, 2006 were analyzed. The 105 donor liver grafts were all used for orthotopic liver transplantation.

RESULTSThe warm ischemia time of the graft ranged from 1 to 8.5 min with a mean of 4 min. The time of graft procurement ranged from 19 to 28 min (mean 22.5 min). Donor liver preparation lasted for 38 to 102 min in the 105 cases, with a mean of 51 min. The cold ischemia time of the donor liver was 5.5 to 13 h (mean 8 h). Anatomical variations were identified in 8 of the donor liver grafts.

CONCLUSIONSCold perfusion of the donor liver and repair of the hepatic artery are important procedures in donor liver procurement and preparation. Hemorrhage due to the donor graft should be prevented and the procedures should be performed in close cooperation with the recipient operation.

Adult ; Female ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Organ Preservation ; methods ; Tissue Donors ; Tissue and Organ Procurement ; methods ; Young Adult

In order to explore the method to prepare hypoxia UW solution and the stability and preservation of hypoxia UW solution, UW solution was purged by argon or air for 15 min or 60 at a flow rate of 0.8 or 2 L/min, and the oxygen partial pressure of UW solution was detected. The hypoxia UW solution was exposed to the air or sealed up to preserve by using different methods, and the changes of oxygen partial pressure was tested. The results showed that oxygen partial presure of 50 mL UW solution, purged by argon for 15 min at a flow rate of 2 L/min, was declined from 242+/-6 mmHg to 83+/-10 mmHg. After exposure to the air, oxygen partial pressure of hypoxia UW solution was gradually increased to 160+/-7 mmHg at 48 h. After sealed up by the centrifuge tube and plastic bad filled with argon, oxygen partial pressure of hypoxia UW solution was stable, about 88+/-13 mmHg at 72 h. It was concluded that oxygen of UW solution could be purged by argon efficiently. Sealed up by the centrifuge tube and plastic bag filled with argon, oxygen partial pressure of UW solution could be stabilized.
Adenosine/chemical synthesis ; Allopurinol/chemical synthesis ; Anoxia ; Glutathione/chemical synthesis ; Insulin/chemical synthesis ; Organ Preservation/*methods ; Organ Preservation Solutions/*chemical synthesis ; Oxygen/*analysis ; Partial Pressure ; Raffinose/chemical synthesis

AIMThe study was designed to examine the effects of cryoprotective media, and glycerolating and thawing procedures on human sperm motility and gel penetrating ability.

METHODFifteen unselected donors provided semen varying in quality that was distributed in a factorial design across three cryoprotectants (glycerol, egg yolk-citrate-glucose-glycerol and egg yolk-tris-glucose-glycerol). Also, glycerol was added at room temperature versus at 4 degrees C. Two thaw temperatures were tested (laboratory air temperature for 10 min versus a 65 degrees C waterbath for 4 seconds). The proportion of total and progressively motile sperm was estimated immediately after thawing and following incubation at 35 degrees C for 2 h. Migration of sperm for 30 min at 37 degrees C through polyacrylamide gel was tested.

RESULTSDonors differed greatly, with post-thaw total motility of sperm ranging from 9 to 44% (P<0.05). Egg yolk-citrate-glucose-glycerol and egg yolk-tris-glucose-glycerol were superior to glycerol alone (post-thaw values of 35, 37 and 21%, respectively, P<0.05). This was due primarily to poor sperm survival when semen was cooled to 4 degrees C without glycerol or egg yolk. The two thaw temperatures gave similar results. Sperm migration tests paralleled the motility results, but were more sensitive in detecting differences.

CONCLUSIONEgg yolk, particularly in a tris-based medium that is widely used in domestic animals, improved the cryopreservation of both good and poor quality human semen.

Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Ejaculation ; Glycerol ; pharmacology ; Humans ; Male ; Organ Preservation Solutions ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; drug effects ; physiology ; Tissue Donors

The effects of oxygen partial pressure on cryopreservation of the cells with organ preservation solution were explored. Hypoxic UW solution was made by purging the UW solution with argon. The pig proximal tubule epithelial cells (LLC-PK1 cells) were cryopreserved in hypoxic UW solution (Ar-UW group) or standard UW solution (UW group) at 4 degrees C for 48 h. Trypan blue staining and LDH detection were performed to evaluate the injury of the cells. The results showed that the oxygen partial pressure in Ar-UW group was significantly declined from 242+/-6 mmHg to 83+/-10 mmHg. After cryopreservation at 4 degrees C for 48 h, LDH leakage rate and Trypan blue-stained rate in Ar-UW group were (11.3+/-3.4)% and (10.5+/-4.7)%, respectively, which were significantly lower than in UW group [(49.5+/-6.9)% and (47.6+/-9.3)% respectively, both P<0.01]. It was concluded that lower oxygen partial pressure of UW solution was more beneficial to the cryopreservation of LLC.
Adenosine ; Allopurinol ; Cell Hypoxia ; Cell Line ; Cryopreservation ; Cryoprotective Agents ; Epithelial Cells/*cytology ; Glutathione ; Insulin ; Kidney Tubules, Proximal/cytology ; Organ Preservation Solutions ; Oxygen/pharmacology ; Raffinose ; Swine ; Tissue Preservation/methods