JPH0822801B2 - Solution for organ preservation - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a solution for organ preservation.

(Prior arts and problems to be solved by the invention) Preservation of kidneys and in vitro storage of cadaveric kidneys are relatively new fields. Preservation of cadaver kidneys for transplantation is commonly performed in hospitals, but progress has been limited to trial and error experiments. Although this study was partially successful from a clinical point of view,
The true atoms behind these successes are not well understood.

As renal transplantation has evolved from rigorous research maneuvers to become an established clinical therapy for end-stage renal disease, renal preservation has progressed from laboratory research stages to established clinical methods. At present, the two most commonly used methods for kidney preservation are the simple hypothermic storage method and continuous perfusion. Simple cryopreservation, the most common method of clinical kidney preservation, removes organs from cadaver donors and chills them. This method is usually performed by a combination of external cooling and short-time perfusion in order to lower the core temperature as quickly as possible. The kidneys are then stored and flushed out in a simple plastic container.
h-out) soaking the container in ice and
Hold at a temperature of ° C. The advantage of this method is its simplicity,
Low cost and ease of internal transplantation. Extensive research has been conducted on the composition of flushing-out solutions for optimal preservation.

The second method of renal preservation that has undergone extensive laboratory research in conjunction with clinical trials is continuous pulsatile perfusion. The basic components of the continuous perfusion method are (1) pulsatile flow, (2) low temperature), (3) membrane oxygenation and (4) a perfusate containing both albumin and lipids. With some variations, all renal preservation units currently in use share these basic principles. The continuous perfusion method in clinical transplantation has several advantages. The primary advantage of perfusion is that there is sufficient time to partially manipulate the transplant from the cadaver. The second is to enable pre-transplant viability testing. If the storage time can be extended to the 5-7 days required for current mixed lymphocyte culture studies, significant improvements in the outcome of kidney transplantation from the cadaver can be expected.

Simple cold storage after flushing with the first intracellular electrolyte solution or electrolyte-protein solution to allow successful preservation of the human kidney by pulsatile perfusion for 2-3 days, a tissue contrast test of donors and recipients, Sufficient time will be available for renal distribution between transplant centers and careful preparatory surgery preparations of recipients, resulting in preliminary donor culture time and pre-transplant donor kidney vessels. You can get repair time. Proving that kidneys stored for 72 hours using cold pulsation perfusion with cold-precipitated plasma represent an important advance in human kidney preservation,
It was the preferred method of preservation. Preservation of renal organs by simple cold storage after treatment with ice-cold intracellular electrolyte flushing solution has been satisfactorily used for human kidney preservation up to 61 hours.

Serum albumin is widely used in clinical organ preservation because it produces the necessary colloid osmotic pressure in various forms. Forms of serum albumin include cryoprecipitated plasma, plasma protein fractions, human serum albumin and silica gel treated plasma. However, since these perfusates are prepared from naturally derived substances, variability is unavoidable. It would be particularly advantageous if a perfusate containing synthetic colloids was available.

In the past, a number of synthetic colloidal substances have been experimentally tested for efficacy in renal preservation. These colloids include dextran, polyvinylpyrrolidone,
Pluronics, hydroxyethyl starch (HES), Ficoll, gum arabic, polyethylene glycol are included. None of these were as effective as serum albumin. However, HES was effective for 24 hours and in some cases 72 hours. All of these colloidal substances were tested in saline-based perfusates. Recently, it has been observed that dog kidneys can be successfully preserved for 72 hours with perfusate containing gluconate anion instead of chlorine in human serum albumin (HSA) for supporting colloid penetration.

There were two important studies in the late 1960s: 30 hours (1) by cold storage of the kidney and 72 by continuous perfusion.
It was shown that it could be safely stored for a long time (2). These two studies changed the clinical transplantation of cadaver kidneys from an emergency procedure to a semi-selective procedure. Many investigators have tested other cold storage solutions (3) and some claimed successful storage for 48 or 72 hours (3-5). However, Collins solution or modified Eurocolli
ns) solution (5) is preferred in most transplant centers where the kidneys are stored by cold storage.

In the 1980s, cyclospoline (cyclospo) for immunosuppression
rine) has revived interest in transplanting other organs, especially the liver, pancreas, heart, lungs, and cardiopulmonary system.
However, successful preservation methods for the kidney have proven unsuccessful for these other organs. Therefore, the heart,
Clinical preservation of the liver and pancreas was kept to a minimum, no longer than 6-10 hours. Transplantation of these organs is several times more complicated than in the case of kidneys, and surgery is always performed at night in the donor's hospital, when the operating room is available. The short shelf life of the heart and liver also requires two surgical teams, one for the donor and one for the recipient. Extending the storage period of these organs to 30 hours has the same impact on transplantation of these organs as that of kidney transplants: increased organ availability, reduced organ disposal, increased organ allocation and reduced costs. Will bring.

Preservative solutions that can be used for both all donor organs, in-situ organ cooling in the donor body and cold storage after organ removal are desirable.

(Means for Solving the Problems) According to the present invention, a perfusate or a storage solution containing a special synthetic HES instead of human serum albumin and an organ preservation method using the same are disclosed. Suitable compositions are provided. Disclosed are storage solutions and perfusates provided for 72 hour preservation of pancreas, 48 hour preservation of kidney and at least 24 hour preservation of liver.

As mentioned above, serum albumin (HSA) based perfusate has been the standard for both experimental and clinical renal preservation for the last 17 years. Unfortunately, this type of perfusate provided a shelf life of only 3 days. Both of these methods preserve the vital activity of the kidney for up to 3 days, but it is difficult to consistently obtain longer preservation times.
In addition, these methods retain viability for up to 3 days, but as shown by increased levels of serum creatinine after kidney transplantation, increasing the time required to restore these elevated levels to normal. The kidney is damaged. The initial perfusate was chosen from the electrolytes readily available for intravenous infusion and was basically of extracellular composition.

To date, no acceptable method has been obtained for renal preservation. Methods that have proven clinically effective are limited to short-term storage (3 days) and viability is greatly diminished. The present invention describes a biosynthetic composition of perfusate and storage solution most suitable for hypothermic organs and a novel synthetic colloid penetrant that results in greatly improved long-term preservation.

Freezing and continuous aerobic perfusion methods are logically true for long term (1
It is the only way to get a preservation for a few years from the month. Simple cold storage has a time limit of the property that the organ loses vital activity. Hypothermia reduces the rate at which intracellular enzymes degrade the major cellular components required for organ vital activity.
Hypothermia does not stop metabolism, it merely slows down kinetics and cell death.

Karn et al., British Medical Journal
1963; 2: 651-655 showed that simple cooling of ischemic kidneys using cold blood retained function for 12 hours. Collins (Lancet 1969; 2: 1219-1222) showed that the use of an appropriate overflow solution increased kidney storage time to function 3 (up to 30 hours). It is considered that this solution was not effective for the preservation of other organs such as the pancreas, liver and heart because of the peculiar difference of organs.

In order for the flash-out solution to be appropriate and effective, 1) minimize cell swelling caused by low temperature,
2) Prevent intracellular acid degradation, 3) Prevent expansion of extracellular space during flash-out period, 4) Prevent damage from oxygen-free radicals especially during reperfusion, 5) During reperfusion Must have a composition that includes a substrate for regenerating the high energy phosphating compounds of Cell swelling caused by low temperature is due to accumulation of water. This swelling tendency can be counteracted by adding a substrate (impermeable agent) that is impermeable to cells (110 to 140 mmol (mmol)) (110 to 140 milliosmol (mOsm) / kg osmotic pressure). The concentration of this impermeant is equal to the concentration of glucose in the cold stock solution of choline (120 mmol) and the concentration of the impermeable agent in the other cold stock solutions. Thus, a key component of a successful cold storage solution is the proper concentration of effective impermeable agent.

A second important consideration for successful cold storage solutions is the prevention of intracellular acid degradation. Ischemia stimulates glycolysis and glycogenolysis (Pasteur effect) even at low temperatures, and also increases lactate production and hydrogen ion concentration. Acidolysis of tissues is crucial to cells, causing destabilization of lysosomal, activating lysosomal enzymes and altering mitochondrial properties. Therefore, prevention of intracellular acid degradation is a prerequisite for good preservation. In one study, an effective buffering or alkaline pH of cold storage solutions was used.
Use of a flash-out solution having
TS et al., Transplant Pros 1984, 16: 134.
-137) and pancreas (Abstract, American Society of Transplant Surgeons,
13th Annual Meeting, May 28-29, 1987).

Effective flash-out solution swells extracellular space,
Swelling that occurs during in-situ flushing of the donor organ and after removal of the organ must be prevented. Such swelling puts pressure on the capillary system and results in poor distribution of the flash-out solution into the tissue. Most cold stock solutions do not contain a substrate on which the colloid-permeants (albumin and other colloids) act. Therefore,
The components of the flash-out solution rapidly diffuse into the extracellular space, causing tissue edema. Therefore, the ideal flash-out solution in that case should contain a substrate that creates a colloid osmotic pressure, with the major components of the flash-out solution freely exchangeable without swelling the extracellular space. .

A fourth important consideration as an effective cold storage solution is the harm of oxygen-free radicals during reperfusion, but the exact role of these agents is also unknown. Endogenous saquintin oxidase has low activity compared to the high endogenous activity of superoxide dismutase, which traps superoxide anions, so oxygen-free radicals are less important for human liver and kidney. Is not considered. In contrast, the harm caused by oxygen-free radicals is extremely important for the lungs and intestines, which are sensitive to such damage.

The last important point to consider is energy metabolism.
Adenosine triphosphate (ATP) decomposes rapidly during cold storage, which results in the formation of end products (adenosine, inosine and hypoxanthine) that can freely penetrate the plasma membrane. Reperfusion of organs requires rapid regeneration of ATP-required sodium pump activity. Therefore, availability of ATP precursors may be important for effective organ preservation.

There are important differences in the metabolism of the kidney, liver and pancreas, which influence how well these organs are preserved. An effective impermeable agent is required to suppress cell swelling. Glucose, the major impermeant of Corinth's solution, is not effective in the liver or pancreas and easily enters cells. Southard et al., Cryobiology 1986; 23: 47.
7-482. Another commonly used impermeable agent, mannitol, penetrates in the liver like glucose. Thus, one of the reasons that cold storage solutions that rely on glucose or mannitol are not effective on the liver and pancreas is that they do not contain effective impermeable agents.

According to the present invention, the organ preservation solution contains lactobionate anion and raffinose as an impermeable agent for cells, a solution osmolality of about 320 million osmoles (mOsm) / l, a solution osmolality of 120 mM, and K ⁺ of 30 mM. Na ⁺
have. Preferred colloids are about 150,000 to about 35
A modified hydroxyethyl starch having an average weight molecular weight of 0,000 daltons and a degree of substitution of from about 0.4 to about 0.7. More preferred colloids are about 200,000 to about 300,000
Hydroxyethyl starch having an average weight molecular weight of Daltons. Preferred colloids are substantially free of hydroxyethyl starch having a molecular weight of less than about 50,000 daltons. According to one embodiment of the invention, the hydroxyethyl starch is dialyzed or treated with distilled deionized water to remove some previously unknown impurities that have an adverse effect on the effectiveness of the hydroxyethyl starch product. The material removed by dialysis is a very small amount of hydroxyethyl starch, which contains residual acetone and sodium chloride along with the by-products of hydroxyethylation, ethylene glycol and ethylene chlorohydrin. Ethylene glycol and ethylene chlorohydrin are known to be toxic. Therefore their removal, even in small amounts, is desirable.

In preferred embodiments, the preservative and perfusate compositions include, but are not limited to:

If necessary, the following additives are added aseptically prior to use to obtain the final solution composition.

1. Penicillin G 200,000 units 2. Regular insulin 40 units 3. Dexamethasone 16 mg For concentration range, the following composition is preferred: hydroxyethyl starch 45.0-57.5 g / l lactobionic acid 32.25-41.20 g / l phosphate ion 2.16- 2.76g / l (monobasic potassium phosphate 3.09 to 3.96g / l equivalent) Sulfate ion 0.43 to 0.55g / l (magnesium sulfate heptahydrate 1.10 to 1.41g / l equivalent) Magnesium ion 0.11 to 0.13g / l (Magnesium sulfate heptahydrate 1.12 to 1.32g / l equivalent) Raffinose pentahydrate 16.05 to 20.50g / l Adenosine 1.21 to 1.54g / l Allopurinol 0.12 to 0.16g / l Glutathione 0.83 to 1.06g / l pH 6.8-8.0 The preferred potassium level is about 120 mEQ / l to about 150 mEQ.
/ l, the sodium level is about 20mEQ / l to 30mEQ / l
Is.

If necessary, remove glutathione from the composition (0
%), And may be packaged as a separate additive such as a lyophilized additive for addition as a final solution composition just prior to use.

For a wider concentration range, the following composition would be a suitable solution for organ preservation: hydroxyethyl starch 2-12% lactobion 1-7% raffinose 0.5-3% adenosine 0.1-0.3% allopurinol 0.01-0.3 % Glutathione 0.0-0.3% Potassium About 120-150 mEQ / l Sodium About 20-30 mEQ / l Water for injection 0.5 Preferred colloids for stock solutions are about 200,000 to about 3
Hydroxyethyl having a weight average molecular weight of 50,000 daltons, a degree of substitution of from about 0.4 to about 0.7, substantially free of contaminants including ethylene glycol, ethylene chlorohydrin, acetone and sodium chloride, and less than about 50,000 daltons. It is a hydroxyethyl starch that does not substantially contain starch.

The solution can be used to simple cold flush and store organs including kidneys, liver and pancreas for pre-transplant transfer. Hydroxyethyl starch is about 15
It has a molecular weight of from 0,000 to about 350,000 daltons and is substantially free of ethylene glycol, ethylene chlorodrine, sodium chloride and acetone, hydroxyethyl starch having a molecular weight of less than about 50,000 daltons.

Preferably, the preservation solution is about 2-12% hydroxyethyl starch, about 1-7% lactobionic acid and about 0.5-3.
% Raffinose.

(Example) Next, the present invention will be described in more detail based on examples.

Reference Example Preparation of Hydroxyethyl Starch 100 g of hydroxyethyl starch was dissolved in 1 of distilled deionized water to make a 10% w / w solution. 50,000 of this HES solution
Place in a dialysis bag (34 mm x 18 inches) that separates the molecular weight of daltons and in a container of 10-15 l of distilled deionized water.
It was stirred for 2 hours. Water was changed daily and HES was collected and frozen at -20 ° C until use.

Example 1 72-Hour Preservation of Dog Pancreas Adult mixed-breed dogs weighing 15-25 kg were used in the experiment.

Surgical operation. Anesthesia starts with pentathol
And continued with halothane. The left section of the pancreas (tail) was removed by centerline dissection as described above. Immediately after flushing out the transplanted part (graft) with spleen (control), after 48 hours refrigeration and refrigeration 72
After a lapse of time, they were transplanted into long bone canal (iliac vessel).
The pancreatic tube was left open and the pancreatic fluid was allowed to drip freely into the abdominal cavity. No anticoagulant was used. The right section of pancreas was removed at the time of transplantation.

Experiment record. All dogs received 0.5 g of Mandol IV before extraction and for the first 3 days after transplantation. The dogs were fed standard dog food containing Viokase. These animals were divided into 3 groups. group
1: Organ removal, graft immediately after washing, group 2
(48 hours cold storage), Group 3 (72 hours cold storage. Blood glucose concentration daily for 1 week after transplantation, then 2
It was measured once a week. Intravenous glucose tolerance test (IV
GTT) was performed 24 hours, 2 weeks and 4 weeks after transplantation. Four
Grafts were removed after a week and IVGTT was performed 2-3 days later. IVGTT was injected with glucose (0.5 g / body weight 1 kg), and blood glucose was measured at 1 minute, 5 minutes, 10 minutes, 20 minutes, 30 minutes, 60 minutes, and 90 minutes thereafter. The K value was calculated from the blood glucose concentration obtained from the 5-60 minute measurement (9). Glucose level is more than 150mg% over 2 days, K value is
A value lower than 1.0 was considered as a sign of diabetes.

Save. A comparison of stock solutions is shown in Table 2. After removal, the pancreas is approximately 250-300 ml of flash-out solution with a height of 60 cm
I flushed from. The grafts were immersed in the stock solution in double plastic containers and the containers were placed in an ice water bath.

Statistical processing. Statistical evaluation was performed using the student T test method.
The values obtained are the mean ± SEM.

Results All transplants (grafts) were well perfused immediately after transplantation. In the preserved transplants, intralobular edema (intralobular edema) occurred to some extent at 5 to 10 minutes after perfusion. In all cases the spleen was well perfused. As shown in Table 2, 5 dogs died. Of these, 3 were in the control group and 2 were in group 3 (stored for 72 hours). The cause of death was unrelated to the transplant, and all dogs died with the graft still functioning.
Upon pancreatic function testing, all grafts showed varying degrees of fibrosis as did spleen (even control). Arterial and venous thrombosis was not apparent in any graft.

Post-transplant blood glucose levels and IVGTT results are shown in Table 2 for each animal. Average value of each group examined (+ SE
M) is also shown in Table 2. Group 3 had the highest mean blood glucose level during the first week after transplantation (124 ± 6 mg
%), This value was significantly different (p <0.05) when compared with Group 1 (94 ± 7 mg%) and Group 2 (107 ± 7 mg%). Average K value of 1 days is also 3 (185 ± 0.15
%) Are group 1 (2.44 ± 0.14%) and group 2 (2.
53 ± 0.22%) was significantly lower (p <0.05). In group 3, K value (1.7
± 0.1%) and K value of 28 days (1.61 ± 0.19%) is 1 day
It remained the same as the K value of (P = NS). Group 1
Also, the K value decreased in Group 2 and there was no significant difference between the 3 groups 2 weeks after transplantation. Until the 4th week, the K value of Group 3 was better than that of Group 2, but
It was slightly lower than that of the control group (statistical significance was not shown due to the small number in group 1).

All dogs developed post-pancreatic hyperglycemia (blood glucose concentration was greater than 200 mg%), indicating that the transplanted organs were significantly involved in glucose homeostasis. Long-term survival was observed in 4 animals in group 3. One dog died of pneumonia 7 weeks after transplant but remained euglycemic. Two dogs were sacrificed after 3 and 4 months and one dog was left for 6 months. All dogs were euglycemic with no signs of diabetes.

Example 2 24-Hour Liver Conservation Clinical liver preservation is limited to about 6-10 hours, and increasing it to 24 hours or more would have a significant impact on liver transplantation. Separated and perfused rabbit livers were used for evaluation of storage properties in Corinth's solution, Cambridge Serum Protein Fraction (PPE), Marshall's solution and the solution of the invention (preservation solution) following cold storage. Bile production during normal temperature perfusion of cold-stored liver is a useful parameter of viability,
Bile production rate of rabbit liver stored cold for 24 hours (ml / 100gm
/ hr ± SD) is shown in the table.

Control 5.4 ± 1.7 Cambridge PPE 1.8 ± 0.9 Eurocorins 1.9 ± 1.3 Marshall 3.1 ± 0.5 Storage solution 4.4 ± 0.5 Description Storage solution is cold storage (2-4 ℃) for 24 hours compared to cold storage solution, bile production by normal temperature perfusion Was excellent in terms of.

The final test of a successful preservation solution is the transplant model. Therefore, the stock solution was used in a canine normal liver transplant model.
Perfusion with this solution followed by the livers of 3 dogs
Stored for 24-26 hours. Implantation was done with a combination of cuff and suture techniques. All livers had a pleasing appearance, 3 dogs were awake and agitated within 4 hours of the end of treatment. Platelet counts were normal 6 hours after surgery. Bilirubin and enzyme values for 6 hours and subsequent 7 days are recorded in the table, indicating a rapid recovery of normal liver function. One dog died of intussusception 5 days after surgery.

Example 3 Renal Preservation Based on this experience, the potential utility of the described cold storage (CS) solution for renal preservation was investigated and its effect on renal function after reperfusion was 1) isolated and perfused in dogs. In the kidney model (IPK), 2) dog autotransplant (autotrans)
plat) model.

1) Canine kidneys were stored cold in Eurocollins (EC) or in cold storage solution (CS) as described for 48 hours, respectively. Kidney function was measured at 37 ° C for more than 90 minutes using oxygenated denatured albumin containing Krebs-Henseleit solution during reperfusion of the IPK model. Urine samples were collected and analyzed every 10 minutes. GFR
(Creatini removal), urine / plasma protein (U / P)
And the fraction sodium reabsorption (% Na) were calculated. The results are shown in Table 3 as the average value together with the standard deviation in parentheses.

Both cold storage kidney groups had reduced renal function (compared to control kidneys) upon reperfusion. CS-stored kidneys significantly improved GFR and sodium resorption during IPK compared to FC-stored kidneys. This improvement in function suggests that kidneys stored in CS can recover cold-warm Ishmic damage faster than kidneys stored in EC.

2) The kidneys of 8 dogs stored in CS for 48 hours were sequentially auto-transplanted. Three animals were sacrificed due to technical complexity (arterial infarction, intussusception). Table 4 shows post-transplant serum creatinine (mean ± SD) of the 5 surviving animals.

This study shows that renal function is well preserved when stored cold in CS solution for 48 hours. Therefore, this solution can preserve the kidney, pancreas and liver and can be used for simple cold storage or continuous perfusion.

Example 4 Solution for organ preservation The solution in Table 1 is a pale yellow, clear, sterile, non-pyrogenic solution for simple cold flushing and storage of organs. Solution is about 320 mOsm osmolality calculated, about 2
It has a sodium concentration of 0 mEq / l, a potassium concentration of about 120 mEq / l and a pH of about 7.4 at room temperature. The solution is intended for flushing and cold storage, where organs including kidneys, livers and pancreas are prepared for storage, transport and ultimately transplantation to the recipient (recipient) upon removal from the donor. Solution in ice at about 2-6 ° C (35.6-42.8 °
After pre-cooling to F), it is used to flush the organs from which the chilled solution is to be separated just prior to or immediately after removal from the donor. The solution remains in the organ vasculature during simple cold storage and during transport.

The solution is for cold storage of organs and is not intended for continuous mechanical perfusion. Use of the solution at the recommended temperature effectively cools the organ and reduces its metabolic requirements. Before being connected to an organ, the solution container allows the solution to flow steadily and at least during flushing.
It should be suspended high enough to obtain a flow rate of 30 ml / min. Flushing should be continued until the organs are uniformly pale and the effluent is clear.

Proposed minimum volume In-vivo flush: -Adult 2-4 liters-Infant 50 ml / kg ex-vivo infusion: -Liver (via portal and gallbladder system) -Adult 1200 ml-Infant 50 ml / kg-Pan or kidney-Adult 300- 500 ml-Infants 150-250 ml / kg Aseptically seal the container with additional solution into a container holding the organ. The organ storage container should be kept in a well-insulated transfer container. Use ice around organ storage containers, but not in containers where ice can directly contact the organ.

The solution can be kept in a plastic bag of 1 liter or more, but should be stored at a freezing temperature of 2-8 ° C (35.6-46.6F).

Accordingly, the present invention provides for extended clinical organ preservation time and minimizes the variability resulting from irrigation prepared from naturally derived materials as synthetic colloids.

Although the present invention has been described in detail and with particular reference to particularly preferred embodiments, it will be understood by those of ordinary skill in the art that modifications can be made without departing from the spirit and scope of the invention. Should be.

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Claims (7)

[Claims] 1. After removal from a living or dead donor,
A solution for simple cryopreservation and storage of organs before internal transplantation into recipients, hydroxyethyl starch 50 g / l lactobionic acid 35.83 g / l monobasic potassium phosphate 3.4 g / l magnesium sulfate heptahydrate 1.23g / l Raffinose pentahydrate 17.83g / l Adenosine 1.34g / l Allopurinol 0.136g / l Glutathione 0.922g / l Potassium hydroxide qs (sufficient) Adjusted to sodium hydroxide pH7.4 Including water qs for injection Wherein the hydroxyethyl starch has a weight average molecular weight of about 150,000 to about 350,000 daltons, a degree of substitution of about 0.4 to about 0.7, is free of insulin, bactrim and texamethasone, ethylene glycol, ethylene chlorohydrin, acetone and A solution that is substantially free of contaminants, including sodium chloride. 2. The solution of claim 1 wherein the hydroxyethyl starch is substantially free of hydroxyethyl starch having a molecular weight of less than 50,000 daltons. 3. After removal from a living or dead donor,
A solution for simple cryopreservation and storage of organs before transplantation into recipients, hydroxyethyl starch 45.0 to 57.5 g / l lactobionic acid 32.25 to 41.20 g / l phosphate ion 2.16 to 2.76 g / l sulfate ion 0.43 ~ 0.55 g / l Magnesium ion 0.11 ~ 0.13 g / l Raffinose pentahydrate 16.05 ~ 20.50 g / l Adenosine 1.21 ~ 1.54 g / l Allopurinol 0.12 ~ 0.16 g / l Glutathione 0.83 ~ 1.06 g / l Water for injection qs pH 6.8 to 8.0, wherein the hydroxyethyl starch has a weight average molecular weight of about 150,000 to about 350,000 daltons, a degree of substitution of about 0.4 to about 0.7, and contains ethylene glycol, ethylene chlorohydrin, acetone and sodium chloride. A solution that is substantially free of contaminants. 4. The solution of claim 3 wherein the hydroxyethyl starch is substantially free of hydroxyethyl starch having a molecular weight of less than about 50,000 daltons. 5. After removal from a living or cadaver donor,
A solution for simple cryopreservation and storage of organs before internal transplantation into recipients, which is hydroxyethyl starch 2-12% lactobionic acid 1-7% raffinose 0.5-3% adenosine 0.1-0.3% allopurinol 0.01-0.3% glutathione. 0.0-0.3% potassium 120-150 mEQ / l sodium 20-30 mEQ / l water for injection qs, wherein the hydroxyethyl starch has a weight average molecular weight of about 150,000 to about 350,000 daltons, a degree of substitution of about 0.4 to about 0.7. A solution containing, without insulin, bactrim and texamethasone, and substantially free of contaminants including ethylene glycol, ethylene chlorohydrin, acetone and sodium chloride. 6. The solution according to claim 5, wherein glutathione is packaged as a separate additive and added to the solution immediately before use. 7. The solution of claim 5 wherein the hydroxyethyl starch is substantially free of hydroxyethyl starch having a molecular weight of less than about 50,000 daltons.