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 purposes of initial preservation solution are rapid cooling of the organ and washing out the intravascular components. After the introduction of University of Wisconsin (UW) solution, it has become the standard organ preservation solution in liver transplantation. However, due to several problems such as high viscosity and potassium concentration, let alone its cost, there has been several attempts to use UW solution in combination with other preservation solutions for initial perfusion of the graft during the organ harvest procedure. In order to evaluate the efficacy of Ringers' Lactated (RL) solution as an initial perfusion solution, we performed orthotopic hepatic allografts on dogs. In 4 dogs, UW solution was used as the initial perfusion solution and in the other 4, RL solution was used. After initial perfusion, UW solution was used successively for the preservation of the graft. All harvested grafts were stored in UW solution. Average cold ischemic time was 163.5 minutes for RL group and 159 minutes for UW group. The two groups were compared in terms of hematologic and biochemical markers before and after the transplantation. The extent of graft injury during cold ischemia and after reperfusion was also compared directly under light and electron microscope. There was no difference in cold and warm ischemic time, anhepatic time, and total operation time between the two groups. The groups did not differ in liver function test, complete blood count and coagulation profile. Histologic exams also showed similar changes between the two groups. In conclusion, RL solution can be used as the initial perfusion solution in hepatic transplantation.
Allografts ; Animals ; Biomarkers ; Blood Cell Count ; Cold Ischemia ; Dogs ; Hepatectomy* ; Humans ; Linear Energy Transfer ; Liver Function Tests ; Liver Transplantation* ; Liver* ; Organ Preservation ; Perfusion* ; Potassium ; Reperfusion ; Tissue Donors* ; Transplants ; Viscosity ; Warm Ischemia ; Wisconsin

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 relationship of seasonal variation with pre- and post-thaw semen parameters as well as the cryosurgical of human spermatozoa.

METHODSA total of 6 414 semen samples were collected from 1 135 donors aged 22 - 32 years by Zhejiang Human Sperm Bank, and divided into spring, summer, autumn and winter groups according to the time of collection. All the samples underwent routine seminal analysis, and the sperm parameters were compared between different seasons. The sperm specimens were cryopreserved in aliquots and analyzed after thawing.

RESULTSThe semen volume was (2.92 +/- 1.17) ml in spring, significantly higher than in summer, autumn and winter ([2.71 +/- 1.07 ], [2.74 +/- 1.15] and [2.83 +/- 1.15] ml, P < 0.05). Sperm density was the highest in autumn ([105.60 +/- 39.76] x 10(6)/ml) as compared with the other three seasons ([101.18 +/- 40.16] x 10(6)/ml, [93.54 +/- 35.10] x 10(6)/ml, and [101.29 +/- 38.37] x 10(6)/ml, P < 0.05). The sperm progressive motility was the highest in spring ([58.49 +/- 10.04] %) and the cryosurgical of sperm the lowest in summer, with statistically significant differences from the other groups (P < 0.05).

CONCLUSIONSeasonal variations affect human semen quality and cryosurgical of sperm. The semen volume, the percentage of progressive motile sperm, the cryosurgical of sperm, and the post-thaw density of progressive motile sperm are higher in spring than during the rest of the year.

Adult ; Cryopreservation ; Humans ; Male ; Seasons ; Semen Analysis ; Semen Preservation ; Sperm Banks ; Sperm Count ; Sperm Motility ; Tissue Donors ; Young Adult

OBJECTIVETo evaluate the effects of tannic acid (TA) treatment on the physico-chemical properties of glutaraldehyde (Glut)-fixed bovine jugular vein (BJV).

METHODSFresh BJVs were treated with Glut or Glut/TA, respectively. The shrinkage temperature, resistance to collagenase or elastase digestion, bio-mechanical properties, and molecular structure of these prepared BJVs were evaluated by Fourier transform infrared spectroscopy.

RESULTSTA treatment resulted in higher shrinkage temperature (P < 0.01), higher resistance to collagenase or elastase digestion (P < 0.01), slightly increased tensile strength (P < 0.01), and elongation at break (P < 0.05) in Glut/TA BJV walls when compared with those of Glut group. Chemical bonds existed between TA and JBV tissue.

CONCLUSIONTA treatment can effectively improve the physicochemical properties of Glut-fixed BJVs.

Animals ; Cattle ; Chemical Phenomena ; drug effects ; Elasticity ; drug effects ; Glutaral ; pharmacology ; Jugular Veins ; pathology ; Organ Preservation Solutions ; pharmacology ; Tannins ; pharmacology ; Tensile Strength ; drug effects ; Tissue Fixation ; methods ; Tissue Preservation ; methods

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

OBJECTIVETo investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs).

METHODSHuman RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined.

RESULTSThe protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples.

CONCLUSIONFast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.

Erythrocytes ; cytology ; Freeze Drying ; Humans ; Tissue Preservation ; methods

Generally, mammalian cells and living tissues can be cryopreserved in a frozen state at very low temperatures over a long storage term. The survival rate of cell suspensions is often acceptable however, living tissues suffer a variety of injuries. In this paper, it was demonstrated that the addition of polyphenols extracted from green tea to conventional cell culture medium and tissue compatible liquid, can control cell proliferation and also preserve tissues for several months at ordinary room temperature, including such tissues as blood vessels, cartilage, islet cells and corneas. This protocol allows a non-frozen living tissue bank to be established using the preservation fluid described.
Animals ; Flavonoids/*pharmacology ; Humans ; Organ Preservation Solutions/*pharmacology ; Phenols/*pharmacology ; Tea/*chemistry ; *Tissue Banks ; *Tissue Preservation ; *Tissue Transplantation ; Transplantation, Homologous

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

Organ transplantation is the most effective treatment for end stage organ failure,and voluntary donation after citizen's death is the only source of transplanted organ in China.Clinically,transplantation organ protection technology plays a critical role in improving the quality of transplantation organs and the prognosis of recipients.On the basis of domestic and worldwide basic research and clinical practice and according to the Oxford evidence classification and GRADE system,the experts organized by Organ Transplantation Doctor Branch of Chinese Medical Doctor Association,Organ Transplantation Group in Surgery Branch of Chinese Medical Association and China Liver Transplant Registry Scientific Committee had compiled and published the Chinese Expert Consensus on Organ Protection of Transplantation(2016 edition) for liver,kidney,pancreas,small intestine,heart,lung transplantation organs.With the support of China Liver Transplant Registry,National Center for Healthcare Quality Management in Liver Transplant,National Quality Control Center for Donated Organ Procurement,National Clinical Research Center for Orthopedics & Sports Medicine & Rehabilitation and National Trauma Medical Organ Protection Committee and combined with recent domestic and worldwide clinical practice and research progress for organ transplantation and organ protection,the Chinese Expert Consensus on Organ Protection of Transplantation(2022 edition)(hereinafter referred to as the consensus) has been formulated in 2022.This edition of the consensus focuses on updating the technical progress and evidence-based medicine of organ procurement,organ preservation,organ transport,and quality evaluation in clinical practice.Additionally,protection of composite tissue for transplantation,mainly including limb graft,has also been included.The organ protection strategy recommended in this consensus aims to promote scientific and standardize clinical organ transplantation work.
Asians ; Consensus ; Humans ; Organ Preservation ; Organ Transplantation ; Registries ; Tissue and Organ Procurement