• Journal of Chest Surgery
• /
• v.27 no.9
• /
• pp.723-731
• /
• 1994

We have modified the isolated perfused working rabbit lung model [IPWL] by perfusing the isolated lung with a hollow fiber membrane deoxygenator.For assessment the stored lung was ventilated with FIO2 0.4 and perfused with 37$^{\circ}$C deoxygenated circulating blood at a rate 5ml/kg/min for several hours until lung failure.We chose to compare our developing solution which contained low potassium and pentastarch with the modified Euro-Collins solution .Experiments were divided into four groups[n=6] based on the type of flushing preservation solution and preservation time.The flushed lungs were then preserved into same solution at 8~10$^{\circ}$C with 100% O2 inflated condition for 1 or 20 hours.These following results were obtained.The IPWL model requires only one animal per experiment and allows for the continuous assessment of aerodynamic performance. This should therefore be used as screening test in lung preservation.One hour preservation groups, there were no significant difference in recovery rates of PaO2, PAP and Paw. Survival time in the one hour preservation groups were very significant long in the Group II[LPPS, p<0.01]. Twenty hours preservation groups, there were no significant difference in the recovery rates of PAP and Paw between Group III[m-ECS] and Group IV[NS], but PaO2 was significantly worse at onset of reperfusion in Group III when compared with Group IV [p<0.05]. And also survival time in the 20 hours preservation groups were significant long in the Group IV [p<0.05].

• Journal of Chest Surgery
• /
• v.30 no.1
• /
• pp.1-10
• /
• 1997

The successful cardiac transplantation depends partly on the donor heart preservation by a solution that will ensure recovery of myocardial function. The purpose of this study was to perform the evaluation of various preservation solutions and to accumulate the data on the requisites for ideal preservation solution. The experimental setup was the constant pressure Langendorffs perfusion system. Isolated rabbit hearts were perfused for 20minutes with unarm Krebs-Henseleit solution, stored for 4 hours in cold preservation solution after cardioplegia, and then were reperfused for 20minutes. The 4 experimental groups were prepared Hartmann's solution group (group 1, control), modified Euro-collins solution group(group II. MEC), modified University of Wisconsin group (group n, MUW), and CK solution(made by the author) group (group W, CK). The parameters for assessing the preservation ability were levels of enzymes in freezed myocardial tissues (lactate, creatine kinase-MB and adenosine deaminase), coronary flow. left ventricular developing pressure and dpldt. In conclusion, the ability of preservation for isolated rabbit heart was excellent in CK solution and modified University of Wisconsin solution, and poor in modified Euro-collins solution, compared with Hartmann solution. CK solution has low potassium concentrations(34.2mEq/L) and includes various substrates to be salutary on myocardial preservation. This fact may indicates the necessity of further refinements in selection or composition of electrolytes and substrates.

• KSBB Journal
• /
• v.24 no.3
• /
• pp.312-320
• /
• 2009

Stable cell preservation is an essential factor in the regenerative medicine for cell therapies and transplantation of biologic materials. In this study, we studied to provide more stable hypothermic preservation by protection of cell damage during the preservation at $4^{\circ}C$. The result of searching for key components that have excellent efficacy in hypothermic preservation of cells, we have identified the fact that the hypothermic preservation adding protein hydrolysates such as yeast hydrolysate is far superior to others. All protein hydrolysates that are derived from animal, plant and microbe sources have superior efficacy, especially the peptides which have molecular weights under 10 kDa have the best efficacy among the components of protein hydrolysate. The protein hydrolysates prevented the decrease of ATP level in the cells caused by hypothermic environment and they inhibited the generation of ROS. Adding antioxidants and control agents of osmotic pressure were showed to have more superior efficacy in hypothermic preservation. Finally, KUL261 solution (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%), the preservation solution developed in this study, showed the best efficacy in both cell viability and cell growth more than other conventional preservation solutions. In conclusion, the improved hypothermic preservation solution that contains the protein hydrolysates as a key component provide the best preservation efficacy. It provides better efficacy than other preservation solutions and will contribute to both the development of regenerative medicine and global commercialization in this therapeutic field.

• Journal of Chest Surgery
• /
• v.33 no.5
• /
• pp.377-384
• /
• 2000

Background: Numerous studies of safe, long term preservation for lung transplantation have been performed using ex vivo models or in vivo single lung transplantation models. However, a safe preservation time which is applicable for clinical use is difficult to determine. We prepared LPDG solution for lung preservation study. In this study we examined the efficacy of LPDG(low potassium dextran glucose) solution in 24-hour lung preservation by using a sequential bilateral canine lung allotransplant model. Material and Method: Seven bilateral lung transplant procedures were performed using weight-matched pairs(24 to 25kg) of adult mongrel dogs. The donor lungs were flushed with LPDG solution and maintained hyperinflated with 100% oxygen at 1$0^{\circ}C$ for a planned ischemic time of 24 hours for the lung implanted first. After sequential bilateral lung transplantation, dogs were maintained on ventilators for 3 hours: arterial resistance were determined if the recipients hourly after bilateral reperfusion and compared with pretransplant-recipient values, which were used as controls. After 2hours of reperfusion, the chest X-ray, computed tomogram and lung perfusion scan were performed for assessmint of early graft lung function. Pathological examinations for ultrastructural findings of alveolar structure and endothelial structure of pulmonary artery were performed. Result: Five of seven experiments successfully finished the whole assessments after bilateral reperfusion for three hours. Arterial oxygen tension in the recipients was markedly decrased in immediate reperfusion period but gradually recovered after reperfusion for three hours. The pulmonary artery and pulmonary vascular resistance showed singificant elevation(p<0.05 versus control values) but also recovered after reperfusion for three hours(p<0.05 versus immediate period value). The ultrastructural findings of alveolar structure and endothelial structure of pulmonary artery showed reversible mild injury in 24 hours of lung perservation and reperfusion. Conclusion : This study suggests that LPDG solution provides excellent preservation in a canine model in which the dog is completely dependent on the function of the transplanted lung.

• Journal of Veterinary Clinics
• /
• v.26 no.5
• /
• pp.445-449
• /
• 2009

For human organ transplantations, histidine-tryptophan-ketoglutarate solution (HTKS) and University of Wisconsin solution (UWS) have been shown to engender similar outcomes as gold standard cold preservation solutions ($4^{\circ}C$). To select the effective preservation solution for cold storage of kidney xenografts in miniature pig, which could be a potential source animal of bio-organs, this study compared early histopathological outcomes of cold preservation injury using HTKS and UWS. Twelve miniature pigs weighing 25.6 to 34.7 kg were divided into two groups (n = 6 per group), UWS group and HTKS group. The kidneys in each group were harvested, cold flushed, and preserved for 0, 24, 48, and 72 hrs at $4^{\circ}C$ with UWS or HTKS, respectively. Histolopathological examinations were assessed on kidney biopsy specimens, taken after each cold storage. The degree of renal injury was scored using 5 different criteria (pyknotic nuclei, disruption of cytoplasm, detachment of epithelium, loss of microvilli, tubular necrosis and loss of glomerular tufts) of the cellular components of the tissue. The degree of kidney damage was increased with prolonged cold ischemia time. UWS and HTKS have at least similar efficacy in kidney preservation within 24 hrs cold preservation time. However, in HTKS group cold-induced injury started to be observed more than in UWS group after 48 hrs of cold storage. In conclusion, UWS and HTKS were equally effective for cold preservation of miniature pig kidney in early preservation times; however, UWS may be more effective at longer preservation times as compared to HTKS.

• KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
• /
• v.7 no.1
• /
• pp.25-30
• /
• 2001

The purpose of this study was to examine how can we keep the freshness of peeled chestnuts in distribution process. The 30 minutes impregnation treatment at the preservation solution of 0.3% consistency showed a good result. The solution treatment prevented from surface yellowing and quality degradation of peeled chestnuts. And, in vaccum packaging after treating chemicals surface drying was faster than the other packaging from 7th day. A vitamin C treatment was no effective to restrain the growth of microbials.

• Journal of Chest Surgery
• /
• v.29 no.10
• /
• pp.1066-1075
• /
• 1996

To compare the efficacy of cardiac preservation, we examined purine metabolites during 24 hours of cold storage($0^{\circ}C$) of the Korean ongrel dog hearts after using three different types of cardioplegic solutions. The hypothermic arrest with total cardiopulmonAry bypass method was employed in 51. Thomas solution(575) and blood cardioplegic solution(BCPS) preservation cases. Specimens were analyzed for levels of adenine nucleotides and their precursors by high performance liquid chromatography. The ATP content in the UW(University of Wisconsin) solution group tends to be higher than that of the combined hypothermic arrest group(575 and BCPS groups) after 2,4,8, and 12 hours of preservation respectively, but there were no significant differences between 575 and BCPS groups. The ADP contents in the UWS and BCPS groups were higher than that of the 575 group at 4,8, 12, and 24 hours, but the difference was not statistically significant between UWS and BCPS groups. The AMP contents did not change significantly in the three groups. The adenosine, Inosine, and hypoxanthine concentrations increased progressively, but the lev l of xanthine was very low in the three groups.

• Journal of Chest Surgery
• /
• v.29 no.8
• /
• pp.816-821
• /
• 1996

Lung transplantation is the established treatment for the end stage lung disedse find preservation of the organ is a major obstacle In performing lung transplantation. For solving this problem, we evaluated the histopathologic changes for various preservation solutions. Male mongrel dogs of similar size and weight (15∼20 kg) were used. The dog lungs were flushed with 4fl normal saline(group 1 'n:5): Modified Euro-Collins solution(group 2 n:5) and University of Wisconsin solution (group 3 : n=6), 60m11kg through a catheter placed in the main pulmonary artery aft r flushing of PGE 1 (20ng1kg). The lungs were preserved for 60 hours and measured dry and wet weights. Histologic specimens were taken every 6 hours and %toed for light microscopic evaluation. The edema ratio of the lungs peaked in 12 hours although there was no difference between the groups. Histologically, alveolar septal changes developed in one case (20%) after 1 hour preservation with normal saline. In case of the University of Wisconsin solution, the alveolar septal distortions and swellings were seen in 1 cases (20%) after 6 hours preservation compared with 3 cases (60%) after 6 hours preservation with Modified Euro-Collins solution. Changes of the pneumocytes were observed after 24 hours preser- vation in group 1, after 48 hours preservation in group 2 and after 60 hours preservation in group 3. We conclude that University of Wisconsin solution might have a superior preservation effect compare to normal saline and Modified Euro-Collins solutions.

• Journal of Chest Surgery
• /
• v.21 no.3
• /
• pp.425-446
• /
• 1988

After 24 hours of preservation under 15 mmHg perfusion pressure the recovery rates of isolated canine hearts were determined. Preservation was performed in a cold room maintained at 4*C with 4 different types of perfusates bubbled with a mixture of 95% 0y and 5% CO~ using a modified perfusion unit designed in our institute. The perfusates used were as follows; Group 1: Krebs-Henseleit solution, Group 2: Krebs solution added by albumin and PGE1. Group 3: Modified Wicomb*s solution, Group 4: Modified Collin*s solution. The extent of myocardial recovery was evaluated using a modified isolated carmine perfusion model by measuring heart rate, systolic arterial pressure, left atrial pressure[LAP] and cardiac output. In addition to the above hemodynamic parameters, biochemical and enzymatic assays from perfusates and electron microscopic changes of the myocardium were also studied. The results were as follows; 1] The heart recovery rates were 41.6%, 53.4% and 108.9% in groups 1, 2 and 3, respectively, and group 3 elicited the best result[p< 0.001]. The heart beat was never recovered in group 4. 2] Recovered systolic arterial pressures[mmHg] were 63.3% in group 1, 94.9% in group 2 and 94.3% in group 3. 3] LAPs[mmHg] were 20 in group 1, 13.5 in group 2 and 11.2 in group 3, which suggested that the best myocardial preservation was elicited in group 3[p< 0.05]. 4] Cardiac output, the sum of aortic stroke volume and coronary leakage, were 69.1% in group 2, and 90.7% in group 3, but these were not statistically significant[p=0.24]. No aortic stroke output was measured in group 1 and 4. 5] The degree of myocardial edema increase was 17.5` in group 1, 24.6% in group 2, 20.9% in group 3 and 55.3% in group 4. But there were no statistical differences in each group[p= 0.08]. 6] CPK-MB[U/L] levels were increased 750% and 332%[p< 0.05], glucose levels[mg/dl] 60.5% and 78.2% and SGOT[U/L] levels 523% and 333%, in groups 2 and 3, respectively. Biochemical and enzymatic assays could not be performed in group 1 and group 4, because of poor recovery of heart beat. 7] Electron microscopic findings in the myocardium of most groups revealed slight to moderate muscle cell and mitochondrial edema. But all these findings were within the limits of reversible change. From these above results, it is suggested that modified Wicomb*s solution seems to be the most useful physiologic salt solution for preservation of the heart. We propose that after further study and improvement, our portable continuous hypothermic perfusion system will contribute to the development of a better preservation method for donor hearts for human heart transplantation.

• Reproductive and Developmental Biology
• /
• v.36 no.3
• /
• pp.163-166
• /
• 2012

This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.