CN117717062A - Erythrocyte preservation solution and preservation, transportation and use method thereof - Google Patents
Erythrocyte preservation solution and preservation, transportation and use method thereof Download PDFInfo
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Abstract
The invention provides a red blood cell preservation solution and a preservation, transportation and use method thereof, belonging to the technical field of cytoprotection, wherein the red blood cells and the cell preservation solution are mixed to obtain a cell suspension, and the cell suspension is preserved under the condition of-4 to-8 ℃, so that compared with the preservation technology of the red blood cells at the temperature of 4 ℃, the preservation time limit is greatly prolonged; compared with the frozen red blood cell preservation technology, the invention does not generate ice crystal damage and does not generate toxic effects of cells and human bodies; by placing the cell suspension at the temperature of-10 to-14 ℃ to form gel state for transportation, compared with the transportation in the common 4 ℃ and cryopreservation red blood cell technology, the method can effectively avoid damage to red blood cells caused by fluid shearing force during liquid transportation by enabling the cell suspension to be in gel state for transportation, and leads to hemolysis of the red blood cells; the cell preservation solution disclosed by the invention has no cell and human toxicity, and can safely and efficiently realize high-quality preservation of cells.
Description
Technical Field
The invention belongs to the technical field of cytoprotection, and particularly relates to a red blood cell preservation solution and a preservation, transportation and use method thereof.
Background
The red blood cells are taken as important blood component resources, so that the long-term preservation of the red blood cells is of great significance to the application of the red blood cell resources. Cryopreservation and cryopreservation are one of the most commonly used methods for preserving erythrocytes, and are widely used in preserving erythrocytes. At present, a common way of preserving erythrocytes is to use preserving fluid to preserve erythrocytes at 4 ℃ for one month. Another way of preserving erythrocytes is by cryogenic freezing with liquid nitrogen using high concentrations of glycerol to achieve long-term preservation of erythrocytes.
The storage of the red blood cells at 4 ℃ is mainly to maintain the basic life activities of the red blood cells by adding anticoagulant and energy metabolism substrate, and the deep low-temperature freezing storage of the red blood cells by liquid nitrogen adopts a high-concentration osmotic protective agent of glycerol for the freezing storage protection of the cells; the glycerol can penetrate through cell membranes to be combined with intracellular water of red blood cells, so that the number of ice crystals generated in the freezing process is reduced, and meanwhile, the generation of needle-shaped ice crystals is reduced, and the probability that cell membranes and organelle membranes are damaged by the ice crystals is reduced to a certain extent.
However, the traditional red blood cell preservation method has a plurality of problems, the time limit of the 4 ℃ preservation method of the red blood cells is one month, the time is short, and the huge waste of red blood cell resources is caused; the freezing preservation of erythrocytes is unavoidable because of the hemolysis phenomenon caused by freezing damage, and the problems of cytotoxicity, human toxicity and high osmotic pressure caused by high-concentration glycerol can not be directly recycled, complicated elution operation is required to be carried out, and the content of residual glycerol is required to be controlled, so that the application of the frozen erythrocytes is greatly limited. Meanwhile, both the 4 ℃ preservation method and the freezing preservation method of the red blood cells face the problem that the red blood cells are damaged by fluid shearing force during transportation, and the released content increases the risk of clinical use of the red blood cells.
Therefore, both the traditional way of preserving erythrocytes at 4℃and the way of preserving erythrocytes by freezing are unavoidable to limit the preservation and use of erythrocytes.
Disclosure of Invention
In view of the above, an object of the present invention is to provide a red blood cell preservation solution and a method for preserving, transporting and using the same; the preservation method greatly prolongs the preservation time limit to more than 3 months; according to the method, the red blood cells are in a gel state by adjusting the ambient temperature, and the damage of the fluid shear force to the red blood cells during liquid transportation can be effectively avoided by transporting in the gel state, so that the hemolysis of the red blood cells is avoided; meanwhile, the cell preservation solution disclosed by the invention is free of cells and human toxicity, and can safely and efficiently realize high-quality preservation of cells.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a cell preservation solution, which takes water as a solvent and comprises the following components in concentration: 1 to 7 percent (m/v) of dextran, 1 to 5 percent (m/v) of glucose, 0.1 to 0.7 percent (m/v) of polyvinylpyrrolidone, 0.01 to 0.05 percent (m/v) of adenine nucleoside triphosphate, and 0.1 to 1 percent (m/v) of gelatin.
Preferably, the cells are erythrocytes.
Preferably, the components are pharmaceutical grade.
The invention provides a preservation method of red blood cells, which comprises the following steps:
(1) Mixing red blood cells with a cell preservation solution according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) And (3) preserving the erythrocyte suspension at the temperature of-4 to-8 ℃.
Preferably, the storage state of the erythrocytes is liquid.
Preferably, the red blood cell density in the red blood cell suspension is (1-30). Times.10 8 cells/mL。
The invention provides a method for transporting erythrocytes, which comprises the following steps:
(1) Mixing red blood cells with a cell preservation solution according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) The red blood cell suspension is transported at the temperature of-10 to-14 ℃.
Preferably, the red blood cells are in a gel state in transportation.
Preferably, the red blood cell density in the red blood cell suspension is (1-30). Times.10 8 cells/mL。
The invention also provides a using method of the transported red blood cells, and the gelatinous red blood cells are placed at 18-26 ℃ to recover into red blood cell suspension for use.
Compared with the prior art, the invention has the following beneficial effects: (1) The red blood cell preservation solution and the preservation, transportation and use methods thereof provided by the invention can realize the low-temperature non-freezing long-term preservation of red blood cells at the temperature of-4 to-8 ℃, and the freezing point of a preservation system is reduced to below-8 ℃ through the effective combination of the freezing-resistant components, so that the suspension after mixing the cells is still kept in a liquid state at the temperature of-4 to-8 ℃, ice crystals are not generated due to freezing, and the damage of the red blood cells caused by 'ice crystal damage' caused by the traditional preservation mode is avoided. Meanwhile, the metabolism rate of the red blood cells is effectively reduced under the storage temperature condition of-4 to-8 ℃.
(2) Compared with a preservation method at 4 ℃, the method of the invention ensures that the red blood cells are still in a liquid non-frozen state under the low temperature condition of-4 to-8 ℃, and simultaneously the metabolism rate of the cells is further reduced under the condition of relatively low temperature, the consumption rate of energy substances such as adenosine triphosphate is reduced, and the service life of the red blood cells is prolonged, so that the preservation time of the red blood cells is prolonged to more than three months from the existing one month, the preservation time of the red blood cells is greatly prolonged, and the problems of resource waste, application limitation and the like caused by the time limit are also broken; meanwhile, the gelatin component added in the invention can enable the erythrocyte suspension to be in a gel state at the temperature of-10 to-14 ℃, so that the risk of erythrocyte damage caused by fluid shearing force can be avoided during transportation of erythrocytes in the gel state, the activity of the transported erythrocytes is ensured, the damage caused by the transportation damage of erythrocytes is avoided, the release of contents is avoided, and finally, the risks of allergy and the like during clinical reinfusion are caused, thereby greatly improving the safety of clinical application.
(3) Compared with the high-concentration glycerol freezing preservation method, the method is safe and efficient, does not need to add toxic high-concentration permeability protective agent, does not need to carry out complicated elution operation during use, avoids severe storage and transportation conditions and reduces the cost, and meanwhile, due to the design of reducing the freezing point of a preservation system, erythrocytes are not influenced by the generation of ice crystals under the low-temperature condition, and the damage to cells caused by the penetration of the ice crystals through cell membranes is avoided.
(4) The components of the cell preservation solution are all of medicinal grade, and the components not only have the effect of reducing the freezing point of a preservation system, but also can maintain the activity of red blood cells and ensure the function of the red blood cells; furthermore, the components dextran and polyvinylpyrrolidone have the function of replacing blood plasma, and the application is wide, so that the cell preservation solution can be directly and integrally returned for treatment after preserving erythrocytes.
Detailed Description
The invention provides a cell preservation solution, which takes water as a solvent and comprises the following components in concentration: 1 to 7 percent (m/v) of dextran, 1 to 5 percent (m/v) of glucose, 0.1 to 0.7 percent (m/v) of polyvinylpyrrolidone, 0.01 to 0.05 percent (m/v) of adenine nucleoside triphosphate, and 0.1 to 1 percent (m/v) of gelatin.
In the present invention, the concentration of dextran in the cell preservation solution is 1% to 7% (m/v), preferably 2.5% to 6.5%, and more preferably 5.5%; the polyvinylpyrrolidone concentration is 0.1% -0.7% (m/v), preferably 0.25% -0.6%, and more preferably 0.55%; the dextran and the polyvinylpyrrolidone are used as non-permeable cell protectants, so that the stability of cell membranes can be effectively protected, and meanwhile, the dextran and the polyvinylpyrrolidone are used as a blood volume extender, so that the osmotic pressure of plasma colloid can be effectively improved after reinfusion, the blood pressure is maintained, and the effect of red blood cell reinfusion treatment is further promoted.
In the present invention, the concentration of glucose in the cell preservation solution is 1% to 5% (m/v), preferably 2% to 4.5%, and more preferably 4%; glucose is taken as a nutrition source and is an important substance for maintaining vital activity of cells, and meanwhile, the glucose also plays a role in regulating the osmotic pressure of a preservation system.
In the present invention, the concentration of Adenosine Triphosphate (ATP) in the cell preservation solution is 0.01% to 0.05% (m/v), preferably 0.02% to 0.045%, and more preferably 0.04%; ATP plays an important role in cells, is a main energy supply source in the cells, participates in biochemical reaction, signal transduction, ion balance, regulation and control of a plurality of cell activities, and is not only the basis of cell vital activities but also the guarantee of cell function maintenance.
In the present invention, the concentration of gelatin in the cell preservation solution is 0.1% to 1% (m/v), preferably 0.3% to 0.8%, and more preferably 0.7%; the gelatin has the function of preventing water from forming crystallization to cause cell damage, ensuring that a preservation system is in a gel state at the temperature of minus 10 ℃ to minus 14 ℃, reducing the osmotic pressure difference between the inside and the outside of the cell, preventing the crystallization of the water in the cell and keeping the activity of the red blood cells.
In the present invention, the cells are erythrocytes.
In the present invention, the components are pharmaceutical grade.
In the present invention, the pH of the cell preservation solution is 6.5 to 7.5, preferably 7.0.
In the present invention, the erythrocyte preservation solution is used after being sterilized by filtration, and is filtered by a sterile filter of 0.22 μm.
The invention provides a preservation method of red blood cells, which comprises the following steps:
(1) Mixing red blood cells with a cell preservation solution according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) And (3) preserving the erythrocyte suspension at the temperature of-4 to-8 ℃.
In the present invention, the storage state of the erythrocytes is liquid.
In the present invention, the density of red blood cells in the red blood cell suspension is (1-30). Times.10 8 cells/mL, preferably (5-20). Times.10 8 cells/mL。
In the present invention, the storage temperature after mixing the erythrocytes with the cell preservation solution is-4 to-8 ℃, preferably-6 to-7 ℃.
The invention provides a method for transporting erythrocytes, which comprises the following steps:
(1) Mixing red blood cells with a cell preservation solution according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) The red blood cell suspension is transported at the temperature of-10 to-14 ℃.
In the present invention, the state of transportation of the erythrocytes is gel-like.
In the present invention, the erythrocytes in the erythrocyte suspensionThe density is (1-30) x 10 8 cells/mL, preferably (5-20). Times.10 8 cells/mL。
In the present invention, the transport temperature of the gel state of the erythrocyte suspension is-10 to-14 ℃, preferably-11 to-12 ℃.
The invention also provides a using method of the transported red blood cells, and the gelatinous red blood cells are placed at 18-26 ℃ to recover into red blood cell suspension for use.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The cell preservation solution was prepared with water for injection, wherein the concentration of dextran was 1% (m/v), the concentration of polyvinylpyrrolidone was 0.1% (m/v), the concentration of glucose was 1% (m/v), the concentration of ATP was 0.01% (m/v), and the concentration of gelatin was 0.1% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 0.5:1, transferring to-6 to-8 ℃ for preservation for 3 months, transferring the liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Example 2
The cell preservation solution was prepared with water for injection, in which the concentration of dextran was 2.5% (m/v), the concentration of polyvinylpyrrolidone was 0.25% (m/v), the concentration of glucose was 2% (m/v), the concentration of ATP was 0.02% (m/v), and the concentration of gelatin was 0.3% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 0.7:1, transferring to-6 to-8 ℃ for preservation for 3 months, transferring the liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Example 3
The cell preservation solution was prepared with water for injection, wherein the concentration of dextran was 4% (m/v), the concentration of polyvinylpyrrolidone was 0.4% (m/v), the concentration of glucose was 3% (m/v), the concentration of ATP was 0.03% (m/v), and the concentration of gelatin was 0.5% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 0.9:1, transferring to-6 to-8 ℃ for preservation for 3 months, transferring the liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Example 4
The cell preservation solution was prepared with water for injection, in which the concentration of dextran was 5.5% (m/v), the concentration of polyvinylpyrrolidone was 0.55% (m/v), the concentration of glucose was 4% (m/v), the concentration of ATP was 0.04% (m/v), and the concentration of gelatin was 0.7% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 1:1, transferring to-6 to-8 ℃ for preservation for 3 months after uniform mixing, transferring liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Example 5
The cell preservation solution was prepared with water for injection, in which the concentration of dextran was 7% (m/v), the concentration of polyvinylpyrrolidone was 0.7% (m/v), the concentration of glucose was 5% (m/v), the concentration of ATP was 0.05% (m/v), and the concentration of gelatin was 1% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 1:1, transferring to-6 to-8 ℃ for preservation for 3 months after uniform mixing, transferring liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Comparative example
Cell preservation solution was prepared with water for injection, in which the concentration of dextran was 7% (m/v), the concentration of polyvinylpyrrolidone was 0.55% (m/v), the concentration of glucose was 4% (m/v), and the concentration of ATP was 0.04% (m/v). Filtering (sterile filter pore size of 0.22 μm) for sterilization.
Mixing red blood cells and cell preservation solution according to the volume ratio of 1:1, transferring to-6 to-8 ℃ for preservation for 3 months after uniform mixing, transferring liquid red blood cell suspension to-10 to-12 ℃ to form gel, placing on a shaking table in the environment of-10 to-12 ℃ for simulated transportation for 48 hours, and recovering to liquid state at room temperature (18-26 ℃) to detect the activity of the red blood cells.
Comparative example 1
The red blood cell suspension was mixed with the red blood cell preservation solution (4:1) according to the instructions of use using a commercially available red blood cell preservation solution (Sichuan Naugel Biotechnology Co., ltd.) and red blood cells were preserved for 3 months at 4.+ -. 2 ℃ and were placed on a shaking table in an environment of 4.+ -. 2 ℃ for simulated transportation for 48 hours, and the activity of red blood cells was detected.
Comparative example 2
Freezing to preserve red blood cells with glycerol concentration of 40% (v/v), storing red blood cells under liquid nitrogen (-196 deg.C) for 3 months, thawing, eluting glycerol, placing on shaker at 4deg.C+ -2deg.C for simulated transportation for 48 hr, and detecting activity of red blood cells.
The specific results are shown in the following table:
and (3) detecting the recovery rate of red blood cells: red blood cells before and after preservation were counted separately, and red blood cell recovery = number of red blood cells after preservation/number of red blood cells before preservation ×100%
TABLE 1 erythrocyte recovery results
Free hemoglobin detection: taking a red blood cell suspension sample, centrifuging at 2000r/min for 10 minutes, taking a supernatant, detecting an absorbance OD value at 405nm single wavelength by using a hemoglobin detection kit (chromogenic method/Biyun day), and calculating the free hemoglobin content according to a standard curve manufactured by a standard product.
TABLE 2 free hemoglobin content results
| Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Comparative example | Comparative example 1 | Comparative example 2 |
| 5.2g/L | 2.9g/L | 1.8g/L | 0.7g/L | 1.7g/L | — | 55.4g/L | 4.6g/L |
Erythrocyte membrane toleration 0.45% physiological saline assay: adding a red blood cell suspension sample into 0.45% (m/v) sodium chloride solution, standing at room temperature for 2h, centrifuging for 10 min at 2000r/min, taking a supernatant, detecting an absorbance OD value at 405nm by using a hemoglobin detection kit (chromogenic method/Biyun day), and calculating the free hemoglobin content according to a standard curve manufactured by a standard substance.
TABLE 3 detection results of erythrocyte membrane tolerance to 0.45% physiological saline (based on free hemoglobin content)
| Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Comparative example | Comparative example 1 | Comparative example 2 |
| 5.3g/L | 2.8g/L | 2.2g/L | 0.9g/L | 2.1g/L | — | 58.1g/L | 4.9g/L |
Erythrocyte permeation 5 μm pore size deformability assay: taking a red blood cell suspension sample, enabling red blood cells to pass through a 5 μm aperture by a suction filtration mode, collecting suspension, centrifuging at 2000r/min for 10 minutes, taking supernatant, detecting absorbance OD value at 405nm by using a hemoglobin detection kit (chromogenic method/Biyun sky), and calculating the free hemoglobin content according to a standard curve manufactured by a standard substance.
TABLE 4 detection of the deformation of erythrocytes through a 5 μm pore size (based on the free hemoglobin content)
| Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Comparative example | Comparative example 1 | Comparative example 2 |
| 5.9g/L | 3.5g/L | 2.3g/L | 1.4g/L | 2.7g/L | — | 57.2g/L | 5.5g/L |
From tables 1 to 4, it can be seen that the erythrocyte preservation system of the present invention can effectively maintain the activity of erythrocytes, and the preservation effects of erythrocytes by the proportions of the protective components with different concentrations in each example are different: in the examples, the recovery of the stored red blood cells was highest and the amount of free hemoglobin was lowest compared to example 4, indicating that the integrity of the red blood cells in example 4 was best maintained; in the test experiments of 0.45% physiological saline tolerance and 5 μm pore deformability of the erythrocytes of each example, the erythrocytes of example 4 released the least amount of free protein, which means that the cell membrane of the erythrocytes of example 4 has better toughness, can withstand low osmotic pressure and maintain the integrity of the cell membrane, and at the same time, will not deform and rupture after passing through a 5 μm filter.
In the comparative example, gelatin is not added, and after the red blood cell suspension is transferred to the condition of minus 10 ℃ to minus 14 ℃, ice crystals are generated, so that red blood cells are damaged and hemolyzed, and gelatin is the key for forming gel state of a preservation system.
Compared with the comparative example, the example has better red blood cell preservation effect, especially compared with the comparative example, the commercial red blood cell preservation solution adopted in the comparative example 1 can preserve red blood cells at the temperature of 4+/-2 ℃, the recovery rate of the red blood cells after preserving the red blood cells for 3 months and being transported in a simulating way is far lower than that of the example, and the red blood cells in the comparative example 1 can not maintain the integrity of cell membranes after preserving the red blood cells for 3 months and being transported in a simulating way, a large amount of free hemoglobin is released, and meanwhile, the better cell membrane toughness and deformability can not be maintained, so that the long-term preservation of the red blood cells can not be supported. Although the preservation effect of the freezing method of comparative example 2 is obviously improved compared with that of comparative example 1 in the long-term preservation effect of 3 months, the preservation effect is still inferior to that of most of examples, and it is difficult to avoid that the freezing method of comparative example 2 needs to further elute high-concentration glycerin component, so that not only is complicated elution operation increased, but also the performance of the erythrocytes is further affected in the eluting process, the clinical application is limited, and meanwhile, the damage of the erythrocytes caused by the fluid shear force generated by transporting cell suspension after the frozen erythrocytes elutes the protective agent is unavoidable, so that the activity of the erythrocytes cannot be better maintained compared with the method of the invention.
As can be seen from the above examples, the present invention provides a red blood cell preservation solution and a method for preserving, transporting and using the same, wherein red blood cells are preserved for a long period of more than 3 months at a relatively low temperature of-4 to-8 ℃ and under a non-freezing condition in a manner of lowering the freezing point of a system, and the preservation time period is greatly prolonged compared with a conventional method for preserving for one month at 4+/-2 ℃; meanwhile, the erythrocyte preserving fluid is a medicinal grade component, so that the complicated operation that glycerol elution is required in a frozen erythrocyte method is avoided, and part of the components have the functions of expanding blood volume and maintaining blood pressure, therefore, the erythrocyte preserving fluid can be returned to a patient along with erythrocytes, and the treatment effect is further improved on the basis of ensuring the reinfusion safety. The method has the advantages that the gel state red blood cell transportation system adopted by the method avoids the problem of red blood cell damage caused by fluid shear force generated during transportation of a liquid system, and fully ensures the activity of the transported red blood cells.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A cell preservation solution, which is characterized by taking water as a solvent and comprising the following components in concentration: 1 to 7 percent (m/v) of dextran, 1 to 5 percent (m/v) of glucose, 0.1 to 0.7 percent (m/v) of polyvinylpyrrolidone, 0.01 to 0.05 percent (m/v) of adenine nucleoside triphosphate, and 0.1 to 1 percent (m/v) of gelatin.
2. The cell preservation solution according to claim 1, wherein the cells are erythrocytes.
3. The cell preservation solution according to claim 1, wherein the component is of pharmaceutical grade.
4. A method for preserving red blood cells, comprising the steps of:
(1) Mixing red blood cells with the cell preservation solution according to any one of claims 1-3 according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) And (3) preserving the erythrocyte suspension at the temperature of-4 to-8 ℃.
5. The method according to claim 4, wherein the storage state of the erythrocytes is liquid.
6. The method for preserving erythrocytes according to claim 4 or 5, characterized in that the erythrocyte density in the erythrocyte suspension is (1 to 30) ×10 8 cells/mL。
7. A method of transporting red blood cells, comprising the steps of:
(1) Mixing red blood cells with the cell preservation solution according to any one of claims 1-3 according to a volume ratio of 0.5-1:1 to obtain a red blood cell suspension;
(2) The red blood cell suspension is transported at the temperature of-10 to-14 ℃.
8. The method of claim 7, wherein the red blood cell is in a gel-like state.
9. The method of claim 7 or 8, wherein the red blood cell density in the red blood cell suspension is (1-30) ×10 8 cells/mL。
10. The method of using the transported erythrocytes according to any one of claims 7 to 9, wherein the erythrocytes in the form of gel are recovered to a suspension of erythrocytes at 18 to 26 ℃.
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