Preparation and application of polysaccharide organ preserving fluid

Abstract

The invention provides an organ preservation (Multiplex-Saccharide, MS) solution composed of polysaccharide substances, and an application of the organ preservation (Multiplex-Saccharide, MS) solution composed of polysaccharide substances and the application of the organ preservation (Multiplex-Saccharide, MS) solution. The preservation liquid is suitable for cold preservation of isolated organs, and belongs to the field of abdominal organ function preservation liquid. The preserving fluid mainly comprises the following components in concentration: hydroxyethyl starch (MW130000), cane sugar, sodium gluconate, sulfate, phosphate solution and the like which are added into 1000mL of aqueous solution. The preserving fluid has the effect of cold preservation (0-4 DEG C) for cells and tissues, especially for abdominal visceral organs. Research data show that the preserving fluid has recovery activity after kidney cold preservation for 18 hours and organ reperfusion, and compared with a standard organ preserving fluid UW fluid used clinically at present, organ function recovery after organ transplantation is faster. The MS organ preserving fluid is expected to replace imported organ preserving fluid, provides better protection for donor preservation of clinical organ transplantation, and has broad clinical application prospects.

Description

Technical Field

[0001] The present invention provides an organ preservation solution for maintaining the survival and / or function of an ex vivo organ during in vitro cold preservation, which is suitable for in vitro preservation of various organs and belongs to the field of medical equipment. Background Art

[0002] Organ preservation fluid is an important substance in the process of donor organ acquisition and preservation. Ex vivo organs need to be irrigated and preserved with fluid. Human organ transplantation is the process of implanting organs such as the heart, lungs, liver, kidneys or pancreas with specific functions from donors into the recipient's body to replace their failed organs. During the transportation of the acquired donor organs in vitro, corresponding liquid measures need to be taken to protect and maintain their functional activity to reduce ischemia-reperfusion injury of the organs after transplantation. At present, the most commonly used method in clinical practice is to use organ preservation fluid to cold (0-4°C) preserve the organs. During the preservation process of the organs immersed in the preservation fluid, tissue cells are in a state of ischemia, hypoxia and energy metabolism disorder (cold preservation injury). The organs resume blood perfusion again after transplantation, and this process leads to oxidative damage after organ reperfusion. Therefore, the quality and composition of the organ preservation fluid are particularly critical to the effect of organ cold preservation. Organ preservation fluid usually contains colloids, non-osmotic substances, pH buffers and a small amount of energy substrates, etc., which are used to maintain the stability of the internal environment of the organ and reduce reperfusion injury.

[0003] Among the existing organ preservation solutions, the domestic preservation solution is mainly hypertonic citrate adenine (HC-A) solution. The components of HC-A solution include mannitol as a non-permeable substance to prevent interstitial swelling, citric acid as a buffer, and highly permeable electrolytes to prevent cell edema; its advantage is low viscosity, which is convenient for flushing organs; but since HC-A solution does not contain substances such as colloids and energy, it is not suitable for long-term cold preservation of organs. At present, HC-A solution is used clinically for cold flushing of organs and short-term (1 to 3 hours) static cold preservation.

[0004] There are many imported organ preservation solutions, including the UW solution from the United States, the Collins solution from Sweden, the histidine-tryptophan-ketoglutarate (HTK) solution from Germany, the Biolaso ​​solution from Poland, the ET-Kyoto solution from Tokyo, Japan, etc. The results of multiple clinical studies indicate that the UW solution has the best effect on the long-term cold preservation of liver and kidney.

[0005] UW solution is an organ preservation solution developed in 1987 by a research team led by Professor Belzer of the University of Wisconsin. The main components of UW solution include: hydroxyethyl starch (molecular weight 200,000), raffinose, potassium lactobionate, and a small amount of glutathione, adenosine, allopurinol, insulin and dexamethasone (see Table 1). UW solution can preserve large animal organs for up to 24 hours, which is a breakthrough in the history of static cold preservation of organs. The preservation solution is currently widely used in the long-term cold preservation of donor organs in clinical practice. However, the formula of UW solution is complex, the country has long relied on imports, and the transportation route is difficult. The high price brings a certain burden to the cost of clinical transplant patients.

[0006] Table 1. UW liquid composition

[0007] Element Content( / L) Element Content( / L) HES200 50g KH2PO4 25mM Raffinose 30mM MgSO4 5mM Lactose Potassium Salt 100mM Allopurinol 1mM Adenosine 5mM Glutathione 3mM penicillin 200,000 U / L Dexamethasone 16mg Osmotic pressure 320mosm / L pH 7.4

[0008] Note: HES200, hydroxyethyl starch with molecular weight of 200,000.

[0009] Problem to be solved

[0010] As of June 2023, there are 183 hospitals in China that are qualified to perform human organ (liver, heart and kidney) transplantation, ranking second in the world in terms of organ transplant volume. In the field of clinical transplantation, donor organs need to be irrigated and preserved with organ preservation fluid, and the amount of preservation fluid used is huge. At present, there is a shortage of long-term cold preservation fluid for organ transplantation in China, and a new preservation fluid that replaces UW fluid is the goal pursued by researchers in the field of transplantation.

[0011] UW solution can effectively preserve a variety of organs for a long time and has become the "gold standard" for organ preservation solutions. Combined with current clinical use, some shortcomings of UW solution have been found: First, the molecular weight of 200,000 hydroxyethyl starch and trisaccharide substance raffinose in UW solution have high viscosity, which causes large solution resistance during organ flushing and may cause difficulty in organ lavage; second, UW solution contains a high concentration of potassium lactobionate, which is complex to synthesize and leads to high potassium ion concentration in the preservation solution, which has the effect of causing cardiovascular dysfunction after the transplant recipient is opened; third, active substances such as heparin, penicillin, dexamethasone and insulin added to UW solution need to be added when the organ is used, which increases the difficulty of using and operating the preservation solution. Fourth, the current market price of UW solution is 3,000 yuan / L, which is expensive.

[0012] In addition, as the waiting list of patients in need of organ transplants grows, the time patients have to wait for a donor is gradually extended, and more organs are obtained from marginal and non-heart-beating donors, high-quality organ preservation is essential for successful transplant outcomes. Although the organ preservation solutions currently used in clinical practice, such as HCA solution, UW solution, and HTK solution, are effective for the lavage and cold storage of donor organs, there is still room for improvement.

[0013] Due to the hypothermic (0-4°C) storage of organs, energy metabolism in cells is carried out at a low level. Long-term cold storage process can lead to the accumulation of intracellular metabolites, providing a basis for the formation of peroxides after organ reperfusion, leading to ischemia / reperfusion injury of donor organs. Previous studies suggest that exploring a substance that maintains the energy balance of tissue cells, such as inhibiting the cell mitochondrial respiratory chain during the cold storage of organs, reducing the accumulation of cell metabolites and acidosis during the organ storage process, thereby avoiding tissue damage during organ reperfusion, is an effective measure. This is also the component lacking in UW fluid. The previous study of the present invention suggests that the use of a reasonable ratio of D-glucose and D-deoxyglucose will improve the preservation effect of the organ. At present, this study has not been reported (it is the preliminary result of this study), and its application in the preservation solution is the feature of the present invention. In addition, this study found that gluconic acid and glucose derivatives, as non-cell membrane permeation substances, can effectively regulate osmotic pressure, prevent cell edema, and improve the preservation effect.

[0014] A large number of previous studies have shown that the activity of enzymes involved in cell activity plays a key role in the cold storage-reperfusion injury of organs. Although 90% enzyme activity inhibition can be achieved by lowering the temperature to about 0°C during organ preservation, the metabolites produced by cell metabolism during long-term cold storage promote organ reperfusion injury, which is also a common problem faced by organ preservation fluids. Enzyme activity and its catalytic efficiency are not only related to temperature and pH, but also to ion types. Our results show that the addition of a certain dose of sulfate ions is involved in regulating the functions of molecules and proteins related to ischemic injury, which may lead to a reduction in the incidence of early graft dysfunction and promote better long-term function after transplantation.

[0015] Studies have shown that low temperature storage can cause cold damage to cell membranes. Recent studies have found that glycine derivatives such as trimethylglycine, such as betaine, have anti-cold damage effects; by adding this substance, it can be used to maintain cell membranes against osmotic damage.

[0016] The MS solution of the present invention uses hydroxyethyl starch 130000 to replace hydroxyethyl starch 200000, and sucrose to replace raffinose, thereby improving the viscosity and liquid stability of the preservation solution and reducing the cost of the preservation solution. In addition, sodium salt replaces potassium salt to reduce the potassium ion concentration in the preservation solution and reduce the effect of the preservation solution on the cardiovascular system.

[0017] Therefore, based on the latest research progress and the above problems with current preservation fluids, this research team invented an organ preservation fluid (also called MS fluid), which can effectively protect cell membranes on the one hand, and reduce the price of preservation fluid on the other.

[0018] To this end, we invented this new polysaccharide (MS) preservation fluid for organ lavage and storage, hoping to use it in clinical practice to improve the difficulties faced by clinical transplantation. Summary of the invention

[0019] Based on the current research on organ preservation fluids, and in response to the research results on the deficiencies of various preservation fluids and organ cold preservation damage, this study invented a polysaccharide-based, highly efficient, multifunctional organ preservation fluid (MS fluid for short). MS organ preservation fluid is suitable for long-term cold preservation of abdominal visceral organs, such as the liver, heart, kidneys, and pancreas.

[0020] The preparation of MS solution is as follows:

[0021] Hydroxyethyl starch (130000): 20-60 g / L,

[0022] Sucrose: 10-35mmol;

[0023] Sodium D-gluconate: 30-80 mmol;

[0024] D-glucose: 5~10mmol

[0025] D-deoxyglucose: 5-10 mmol;

[0026] Betaine: 10-50mmol;

[0027] Glycine: 1-10 mmol;

[0028] Sodium sulfate: 10-50 mmol;

[0029] Potassium phosphate: 10-50mmol;

[0030] Magnesium chloride: 1-10 mmol;

[0031] The pH value of the MS solution provided by the present invention is between 7.4 and 7.5, and the osmotic pressure is 320mOsmol / L; the components of the preservation solution have no incompatibility, good solubility, stable ingredients, and long storage time; it is recommended that the prepared finished product be stored at 0 to 4°C for safety. In addition, the components of the MS solution are simple to synthesize, there are many domestic manufacturers, the price is relatively low, and it is suitable for wide promotion.

[0032] The characteristics of MS fluid are

[0033] Hydroxyethyl starch (HES130) with a molecular weight of 130,000 is used as the colloidal osmotic pressure component of the preservation solution to maintain the stability of the organ's microcirculation; D-gluconate is used as a semi-permeable substance to maintain the osmotic pressure of the cells; glycine and triglycine betaine are used as small molecular substances to resist the osmotic damage of the preservation solution and maintain the integrity of the cell membrane; D-deoxyglucose is used to reduce the accumulation of cellular energy metabolism products and maintain low-level cellular energy metabolism; the disaccharide substance sucrose is used as a stabilizer for the preservation solution; sodium sulfate solution is used to stabilize the activity of metabolic-related enzymes during the preservation process and reperfusion; a low-concentration phosphate buffer is used to maintain the pH value during organ preservation.

[0034] The preservation solution we invented has a lower viscosity, making it easier to use and allowing for more rapid and efficient perfusion of the donor organ. These factors have the potential to significantly improve transplant outcomes by reducing initial tissue damage during transplantation.

[0035] MS solution preparation process

[0036] Step 1: Prepare phosphate solution: Add potassium dihydrogen phosphate / dipotassium hydrogen phosphate into a 750ml double distilled water container and stir to fully dissolve;

[0037] Step 2: Prepare hydroxyethyl starch solution: add hydroxyethyl starch to the above double distilled water container, stir to dissolve at room temperature; continue to heat to 60°C to promote the starch to fully stretch;

[0038] Step 3: Add the following substances to the above solution: sucrose, gluconate, D-deoxyglucose and D-glucose, betaine and glycine, stir to fully dissolve;

[0039] Step 4: Continue to add sodium sulfate and magnesium chloride to the above solution; adjust the volume by adding double distilled water until the total volume of the solution is 1000ml, and stir until the solution becomes a uniform, colorless, clear liquid. Use KHO to adjust the pH of the solution to 7.4. After activated carbon filtration, encapsulate the filtrate, sterilize it at high temperature and high pressure (109℃, 45 minutes), and store it at 4℃.

[0040] Beneficial Effects

[0041] Compared with the existing UW organ preservation solution, the MS organ preservation solution provided by the present invention has the following beneficial effects:

[0042] (1) The MS solution provided by the present invention is used for cold perfusion and in vitro cold storage of various organs, and can effectively statically cold preserve the functions of the kidneys, livers and hearts of rats for up to 24 hours.

[0043] (2) Sulfate was used to improve energy metabolism during cold ischemia-reperfusion, and phosphate buffer was used to maintain the pH value in the organ microenvironment. Hydroxyethyl starch with a molecular weight of 1300 was used to effectively maintain the organ's microcirculation system, and sucrose was used to stabilize the preservation solution. This study used gluconic acid and glucose derivatives as non-cell membrane permeable substances, which can effectively regulate osmotic pressure, inhibit cell edema and intercellular edema, and improve the preservation effect.

[0044] (3) The organ preservation solution provided by the present invention contains betaine and glycine, which can safely and effectively reduce the cell membrane permeability damage during the cold storage of organs, maintain the integrity of the cell membrane, and prolong the preservation time due to the inflammatory response caused by the non-physiological state. The glucose derivative 2-deoxyglucose is used to inhibit the sugar metabolism process (glycolysis process) in cells and reduce the accumulation of metabolites, thereby reducing the ROS formation of organs after reperfusion.

[0045] (4) The current clinical organ preservation uses UW solution based on hydroxyethyl starch (HES200), which has high viscosity and affects the perfusion process. Our research shows that replacing HES200 with hydroxyethyl starch with a molecular weight of 130,000 can reduce the damage of donor organs during cold storage. Static cold storage and reperfusion experiments on rat and pig kidneys showed that the use of MS solution can maintain the functional state of the organ with a protective effect similar to that of UW solution, and can replace UW solution in clinical organ transplantation. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solution in the patent embodiment of the present invention, the following is a brief introduction to the drawings required for use in the specific implementation scheme.

[0047] Figure 1. HE staining and sectioning of rat kidneys 24 hours later to examine renal structure; A: Tissue sections after 24 hours of UW liquid-cold lavage and static cold storage showed that the glomeruli and renal tubules were basically normal; B: Kidneys cold-preserved with the MS preservation liquid of the present invention showed that the renal tubules were also basically normal, which was better than that of the UW group.

[0048] Figure 2.The tissue structure changes of the rat kidneys after cold storage for 18 hours / reperfusion for 7 days in the two groups of preservation solutions. The kidneys cold stored for 18 hours were transplanted, and the rats survived for 7 days after the operation. The HE staining sections of the kidneys showed that compared with the kidney structure of the UW group, the renal tubules of the MS group of the present invention were slightly dilated.

[0049] Figure 3. Changes in tissue results after static cold storage of experimental pig kidneys for 24 hours and reperfusion in vitro for 6 hours. HE staining section tissue examination showed that the MS preservation solution (B) of the present invention protected the renal tubular structure more significantly than the renal structure of the UW group, and no obvious epithelial cell shedding and renal tubular necrosis were observed. Specific implementation plan

[0050] The following examples (1-3) and comparative examples (1-6) are used to further illustrate the synergistic effect between the various components of the present invention. The various components of the present invention are all available on the market. The total amount of the preservation solution in each of the following examples is 1 liter:

[0051] Table 2. Composition of the storage solution of each embodiment and comparative example

[0052]

[0053]

[0054] Note: The unit of hydroxyethyl starch (HES130 or 200) is g / L, the unit of osmotic pressure (crystal) is mOsmol / L, and the unit of osmotic pressure (colloid) is mmHg. In Examples 1-3, the preservation solution is prepared by adjusting the content and ratio of each component, while in Comparative Examples 1-6, the preservation solution adds, replaces or deletes important components; wherein Comparative Example 1 uses sodium chloride instead of sodium sulfate on the basis of HES200, and the components in Comparative Example 2 do not contain gluconate but are replaced by glucose and sodium chloride; Comparative Example 3 does not contain glycine and trimethylglycine (betaine), Comparative Example 4 does not contain D-deoxyglucose, Comparative Example 5 does not contain D-deoxyglucose, glycine or betaine, and in Comparative Example 6, none of the three substances gluconate, D-deoxyglucose, glycine and trimethylglycine are included, and in order to achieve consistent overall osmotic pressure, its components are replaced by sodium chloride.

[0055] Specific experiment 1. Study on in vitro cold storage of rat kidney

[0056] Materials and methods: Male SD rats weighing 250-300 g were used. Both donor and recipient rats were SD rats. Rats were anesthetized by intraperitoneal injection of 1% sodium pentobarbital at 10 ml / kg. The skin was prepared and disinfected, and a midline incision was made in the supine position. The skin and muscles were cut along the midline of the abdomen from the xiphoid process to the pubic symphysis to enter the abdominal cavity.

[0057] Obtaining the donor kidney: Expose the left kidney, free the renal artery and renal vein from the proximal end to the distal end to the renal hilum, and ligate the renal vein branch with 8-0 microsutures. Free the ureter to 2 cm below the lower pole of the kidney and cut the ureter. After the renal artery, renal vein, inferior vena cava and abdominal aorta are fully freed, 1 ml of heparinized saline (100U / ml) is injected into the dorsal vein of the penis to heparinize the recipient mouse systemically. Block the abdominal aorta above and below the level of the renal artery with a vascular clamp, puncture with a 28G indwelling needle, and fix the needle. Slowly and evenly inject the organ preservation solution of each group at 4℃, and continue to inject the perfusion solution until the effluent from the vein is colorless and the kidney is pale in color, with a total amount of about 5ml. Cut the renal artery near the abdominal aorta and remove the kidney. The obtained kidney is placed in a 20ml container in a refrigerator at 0-4℃ for corresponding in vitro static cold storage. According to the different preservation solutions used, the experiment was divided into corresponding groups (N=4 in each group). After 24 hours of static cold storage, the kidneys were irrigated from the renal artery and the irrigated fluid was collected for index detection, including the lactate dehydrogenase (LDH) activity of renal tissue damage.

[0058] The experimental results show that among the comparative examples, the results show that the comparative examples 1-2, which simultaneously contain sodium gluconate, betaine, and deoxyglucose, have better effects than comparative examples 3-5 (Table 3), while the comparative example 6 lacking these three components has the worst effect on kidney cold preservation; and the protective effect of Example 1 is better than that of Comparative Examples 1 and 2. Among Examples 1-3, Example 1 has the best effect (Table 4). Therefore, based on the renal transplantation model and the large animal renal perfusion model, we further evaluated the preservation effect of the preservation solution of Example 1 (i.e., MS solution) by comparing it with the current standard UW solution.

[0059] Table 3. LDH enzyme activity levels in renal artery lavage fluid after cold storage in each comparative example (N=4, x±s)

[0060] Table 4. LDH enzyme activity level in renal artery lavage fluid after cold storage in each example (N=4, x±s)

[0061] Specific experiment 2. Rat kidney transplantation study

[0062] Materials and methods: Male SD rats weighing 250-300g were used. Both donor and recipient rats were SD rats. Rats were anesthetized with 1% sodium pentobarbital, 10ml / kg intraperitoneally. Donor surgery and cold storage methods were the same as those of the specific experimental plan. 1. The anesthesia, body position and kidney release of recipient rats were the same as those of donor rats. The animals had bilateral kidneys removed, and the left kidney was used as the transplantation surgery area. The end-to-end anastomosis method was used, and the renal artery and renal vein were anastomosed end-to-end with 10-0 needle and thread. The proximal vascular clamp of the anastomosis was opened, and the color and texture of the kidney after perfusion were observed, and whether there was urine flowing out of the ureteral orifice. The ureter was reconstructed with 10-0 needle and thread, and the abdominal cavity was checked for active bleeding. The abdominal cavity was flushed with warm saline to facilitate the rewarming of the recipient rat. The muscles and skin were sutured with 3-0 needle and thread, and the operation was completed.

[0063] According to the different preservation solutions used, the experimental rats were divided into a control group (UW group) and an experimental group (MS group); the control group used UW solution, and the experimental group used MS solution, N = 10. The body weight of the rats was observed and various indicators were tested after surgery. The degree of renal tubular injury was detected by HE and TUNEL staining, and blood was drawn 1 day and 7 days after reperfusion to detect blood creatinine and urea nitrogen levels to evaluate the degree of renal injury; the protective effect of different organ preservation on renal cold preservation / reperfusion injury was analyzed.

[0064] Experimental results suggest:

[0065] (1) HE staining showed that the MS solution in the experimental group maintained the tissue structure of the kidney during cold storage; compared with the UW solution in the control group, the renal tissue structure of the MS group was significantly superior (e.g. Figure 1 Show).

[0066] (2) The donor kidneys were stored cold for 18 hours. The survival rates 7 days after transplantation were: 15% in the UW group and 55% in the MS group. MS solution was significantly better than UW solution. There was no significant difference in serum creatinine level, a renal function indicator, between the two groups 7 days after surgery. These results suggest that MS solution maintained renal activity and promoted the recovery of renal function 7 days after transplantation (e.g. Figure 2 Show).

[0067] Specific experiment 3. Cold preservation-reperfusion in vitro experiment of pig kidney

[0068] Experimental materials and methods: Bama miniature pigs, male, weighing 50-80 kg, were used. Miniature pigs were anesthetized by intramuscular injection of ketamine (15-20 mg / kg). The abdomen was opened through a midline incision, and the bilateral kidneys were irrigated with 0-4°C saline. After the kidneys turned white, the bilateral kidneys were removed from the junction of the abdominal aorta and the renal artery. Different organ preservation solutions (UW solution or MS solution) were used to irrigate the kidneys in vitro, with an irrigating height of 70 cm and a time of 15 min (6-8 ml / min), and then the irrigated kidneys were statically cold stored.

[0069] Normothermic ex vivo oxygenation perfusion system: After 24 hours of static cold storage, the kidneys were perfused with an ex vivo normothermic oxygenation perfusion system. Indicators of the ex vivo perfusion system: oxygen flow: 90L / min, perfusion pressure 75mmHg, perfusion fluid gas content and composition: the system ventilation routinely uses 95% O2 and 5% CO2 to ensure oxygen saturation and maintain acid-base balance. During the 6-hour normothermic perfusion of the kidneys in the two groups, specimens were obtained to test relevant indicators. The degree of renal injury was assessed by detecting the enzymatic indicator lactate dehydrogenase (LDH) in the perfusion fluid that reflects cell membrane damage; renal tubular function was analyzed by calculating urine production; tissues were obtained for pathological staining to analyze changes in renal tissue structure.

[0070] The kidney tissues of the two groups were stained by HE after static cold storage for 24 hours / perfusion for 6 hours. Compared with the UW group, the optical microscope of the MS preservation solution of the present invention showed normal cell morphology, tight structure between cells, complete glomerular structure, normal morphology of renal tubular epithelial cells (such as Figure 3) After 6 hours of in vitro normothermic perfusion, all functions were intact, and the kidneys produced 15 ml of urine in the first hour and 100 ml in 6 hours, which was significantly higher than the 80 m / h of the UW group.

[0071] In summary, our results show that the Example 1 group (patented MS solution) reduced cold damage and maintained renal tissue structure during long-term cold storage of rat kidneys. After transplantation, organs preserved with MS solution reduced organ ischemia-reperfusion injury, improved graft survival, and prolonged graft survival. The present invention is not limited to cold storage of kidneys, but can also be used for the preservation of other donor organs such as pancreas, liver, heart and lungs.

Claims

1. An in vitro organ preservation solution, characterized in that: Each liter of preservation solution contains the following components: hydroxyethyl starch: 20-60 g / L, sucrose: 10-35 mmol, sodium D-gluconate: 30-80 mmol, D-glucose: 5-10 mmol, D-deoxyglucose: 5-10 mmol, betaine: 10-50 mmol, glycine: 1-10 mmol, potassium phosphate: 10-50 mmol, sodium sulfate: 10-50 mmol, magnesium chloride: 1-10 mmol, and the balance is deionized water.

2. The in vitro organ preservation solution according to claim 1, characterized in that: The concentration of hydroxyethyl starch (MW 130000) contained in the preservation solution is 50 g / L.

3. The in vitro organ preservation solution according to claim 1, characterized in that: The concentration of sodium sulfate contained in the preservation solution is 10 to 50 mmol.

4. The in vitro organ preservation solution according to claim 1, characterized in that: The concentration of D-deoxyglucose is 5 mmol.

5. The in vitro organ preservation solution according to claim 1, characterized in that: The concentration of the sodium gluconate is 30-80 mmol.

6. The in vitro organ preservation solution according to claim 1, characterized in that: The concentrations of glycine and trimethylglycine (betaine) are: 5 / 15 mM.

7. The in vitro organ preservation solution according to claim 1, characterized in that: The preservation solution contains sucrose at a concentration of 10 to 35 mmol.

8. The method for preparing an in vitro organ preservation solution according to any one of claims 1 to 7, characterized in that: The preparation method of the organ preservation solution is as follows: (1) Prepare phosphate solution: add potassium dihydrogen phosphate / dipotassium hydrogen phosphate into a 750 ml double distilled water container, add deionized water and stir to fully dissolve, to obtain solution A; (2) preparing a hydroxyethyl starch solution: adding hydroxyethyl starch to the solution A, stirring at room temperature to dissolve it; continuing to heat to 60° C. to promote full stretching of the starch, thereby obtaining a solution B; (3) adding sucrose, gluconate, D-glucose, D-deoxyglucose, betaine and glycine to the solution B, stirring to fully dissolve them, and obtaining a solution C; (4) Add sodium sulfate to the solution C; add magnesium chloride after it is fully dissolved, and add deionized water until the total volume of the solution is 1000 ml, stir until the solution becomes a uniform, colorless, clear liquid, adjust the pH of the solution to 7.4 with KOH, filter the filtrate with activated carbon, seal it, sterilize it with high temperature and high pressure, and store it at 4°C.

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