Cell protection liquid for venous transfusion as well as preparation method and application of cell protection liquid

Abstract

The invention discloses a cell protection solution for venous transfusion as well as a preparation method and application of the cell protection solution. The cell protection liquid comprises 4 to 8 mg / ml of sodium chloride, 0.1 to 0.5 mg / ml of potassium chloride, 2.8 to 3.5 mg / ml of sodium lactate, 0.1 to 0.3 mg / ml of calcium chloride, 0.15 to 0.3 [mu] g / ml of glucose, 0.01 to 0.03 ml / ml of 20% human serum albumin, 4 to 8 [mu] g / ml of vitamin E, 0.2 to 2 [mu] g / ml of vitamin B12, 0.5 to 2 [mu] g / ml of folic acid, 4 to 8 [mu] g / ml of L-aspartic acid and 0.5 to 2 [mu] g / ml of glutathione. The cell preserving fluid provided by the invention can effectively protect cells for a long time, can be directly transfused back through veins, does not need special treatment, is convenient for clinical application, and has better application prospect and development potential in the development of the clinical treatment of the cells towards the drug direction.

Description

Technical Field

[0001] The embodiments of the present invention relate to the field of biomedical technologies, and particularly to a cell protective solution for intravenous infusion, a preparation method thereof, and an application thereof. Background Art

[0002] Mesenchymal stem cells (MSCs) have the ability to secrete a variety of neurotrophic factors and have the potential for self-renewal, multi-directional differentiation, and immunomodulation. Therefore, mesenchymal stem cell transplantation has great potential in the treatment of various neurological diseases such as ischemic stroke, multiple sclerosis, Parkinson's disease, and spinal cord injury. In particular, it is widely used in biomedical fields such as the nervous system, osteoarticular system, and genetic immune system. Stem cell therapy is a new field. Developing targeted stem cell drugs and targeted treatment programs is the future development direction, and stem cell therapy brings new hope to patients.

[0003] Intravenous infusion of stem cell transplantation is a widely used clinical treatment method and can be used to treat various diseases such as blood system diseases, immune system diseases, and neurological diseases. During stem cell transplantation, choosing a suitable transplantation route is crucial for the treatment effect. And intravenous infusion as a stem cell transplantation has the following many advantages and is one of the most commonly used stem cell transplantation methods in current clinical treatment and research applications.

[0004] 1. Convenient and fast: Intravenous infusion is a very convenient route. Stem cells can be directly delivered into the patient's systemic circulation system through intravenous injection. Compared with other routes, intravenous infusion is more common, simple, and fast, reducing the discomfort of patients and improving the treatment acceptance of patients.

[0005] 2. Improve the systemic utilization rate: Intravenous infusion can directly introduce stem cells into the patient's circulation system, thereby making greater use of the potential of stem cells. Stem cells can reach various tissues and organs through the circulation system in the body and play their therapeutic role to promote tissue repair and regeneration.

[0006] 3. Wide range of applications: Intravenous infusion is applicable to various types of stem cells such as hematopoietic stem cells and mesenchymal stem cells. Whether it is for treating blood system diseases, organ injuries, or immune system diseases, stem cell transplantation can be carried out through the way of intravenous infusion, having a wide application prospect.

[0007] 4. Reduce the risk of infection: As long as skin disinfection is done well, there is no need for special care like during surgery, and it can largely avoid the external contamination of stem cells during the input process, reducing the risk of infection.

[0008] Intravenous reinfusion can effectively reduce the risk of infection, increase effective utilization, reduce discomfort, and improve treatment effects. Compared with other routes, intravenous reinfusion has a wider range of applications and more advantages. It is one of the routes of stem cell transplantation. At present, there are many objective factors that make stem cells inconvenient to use in terms of culture environment, cell culture and cycle, preparation, and implantation. In addition, the construction of cell therapy in various places requires a lot of resources, funds and equipment investment, resulting in high treatment costs, which may be difficult for patients to bear. Moreover, the time cycle for newly cultured stem cells is relatively long, the safety detection time is long, and long-term in vitro residence causes a decrease in cell viability and an increase in cell fragments. Infusion not only reduces the efficacy, but also has adverse effects on the body.

[0009] Existing cell protection solutions are basically divided into cryopreservation type cell protection solutions and low temperature type cell protection solutions.

[0010] 1. Cryopreservation protective solution basically contains DMSO, which is toxic to cells. Centrifugal washing is required before infusion, which causes certain damage and loss to the cells. In addition, the corresponding environment and equipment are required when processing cells, which increases the cost and difficulty of application.

[0011] 2. Most of the existing low-temperature protective solutions have relatively complex components, including pH buffer, antioxidants, multiple osmotic pressure stabilizers, multiple oxygen free radical scavengers, and other components of cell metabolism. They only consider the protection of cells, but ignore the safety and convenience of direct application. Centrifugal cleaning is still required before application, which will also cause certain damage and loss to the cells. In addition, the corresponding environment and equipment are required when processing cells, which increases the cost and difficulty of application. Summary of the invention

[0012] To this end, the embodiment of the present invention provides a cell protection solution for intravenous reinfusion and a preparation method and application thereof. The cell preservation solution provided by the present invention can effectively protect cells, and is safe to use. It can be directly intravenously reinfused without special treatment, which is convenient for clinical application. The development of the cell protection solution by the present invention ensures the quality, effectiveness, safety, convenience, and promotion of cell drugs during application, and has good application prospects and development potential in the development of clinical cell treatment towards pharmaceuticalization.

[0013] In order to achieve the above purpose, the embodiment of the present invention provides the following technical solutions:

[0014] According to the first aspect of the embodiments of the present invention, the present invention provides a cell protective solution for intravenous transfusion, and the cell protective solution includes: sodium chloride 4 - 8 mg / ml, potassium chloride 0.1 - 0.5 mg / ml, sodium lactate 2.8 - 3.5 mg / ml, calcium chloride 0.1 - 0.3 mg / ml, glucose 0.15 - 0.3 μg / ml, 20% human albumin 0.01 - 0.03 ml / ml, vitamin E 4 - 8 μg / ml, vitamin B12 0.2 - 2 μg / ml, folic acid 0.5 - 2 μg / ml, L - aspartic acid 4 - 8 μg / ml, glutathione 0.5 - 2 μg / ml.

[0015] Under normal temperature environment, vigorous cell metabolism will produce a large amount of oxygen free radicals and lipid peroxides, and will also cause changes in environmental pH and osmotic pressure, resulting in cell swelling and death. Under low - temperature conditions, cell metabolism is lower. In addition, the added sodium chloride, potassium chloride, sodium lactate, and calcium chloride can maintain the osmotic pressure balance inside and outside the cells, reduce the damage to cells caused by osmotic pressure changes, and keep the cells in a relatively stable environment, reducing the metabolic stress response to a certain extent.

[0016] As an important energy substance, glucose can provide basic energy supply for cell metabolism, meet the certain nutritional needs of stem cells even in the low - metabolism state, and maintain the basic life activities of cells.

[0017] 20% human albumin can maintain the plasma colloid osmotic pressure, stabilize the liquid environment around cells during cell transfusion, and can also bind and transport some small - molecule substances, supporting the material transport and metabolism of cells. At the same time, it can provide certain structural support for cells and protect cells from mechanical damage.

[0018] Vitamin E is a strong antioxidant, which can effectively scavenge oxygen free radicals generated inside cells, reduce lipid peroxidation reactions, protect the integrity of cell membranes, and reduce the degree of cell damage.

[0019] Glutathione is a natural substance composed of three amino acids. It can scavenge harmful free radicals, help with detoxification, maintain normal cell metabolism, and enable antioxidants such as vitamin C and vitamin E to play a repeated role, protecting cells from damage.

[0020] L - aspartic acid participates in the nitrogen metabolism inside cells, can provide a nitrogen source for cells, and also helps to maintain the acid - base balance inside cells, playing an important role in the normal metabolism and function maintenance of cells.

[0021] Vitamin B12 and folic acid participate in the nucleic acid synthesis and DNA repair processes of cells, which are crucial for cell growth, proliferation, and repair. During intravenous transfusion, they can help cells maintain normal genetic material stability and repair ability, and promote cell survival and functional recovery.

[0022] The cell preservation solution provided by the present invention has relatively simple components, including substances that maintain the stability of the intracellular environment such as electrolytes like sodium chloride and potassium chloride, glucose that provides energy, 20% human albumin that maintains the cell structure and function, as well as vitamin E, B12, folic acid, L-aspartic acid, glutathione, etc. that have antioxidant and nutritional effects. It can not only protect cells but also be directly used for intravenous infusion, improving safety and convenience. Verified by experiments, after being preserved with the cell preservation solution, the cell viability is not affected, and at the same time, the cell morphology is normal, its proliferation and amplification ability is not affected, its phenotypic characteristics are not affected, and it is safe and reliable.

[0023] The cell preservation solution provided by the present invention does not use toxic components such as DMSO, reducing the toxic damage to cells. It also does not require complex centrifugation and washing steps, avoiding cell damage and loss during the washing process, reducing the application cost and difficulty. In addition, the main components are of medical grade and can be directly input into the human body without any risk.

[0024] Further, the cell preservation solution includes: sodium chloride 6 mg / ml, potassium chloride 0.3 mg / ml, sodium lactate 3.1 mg / ml, calcium chloride 0.2 mg / ml, glucose 0.23 μg / ml, 20% human albumin 0.02 ml / ml, vitamin E 6 μg / ml, vitamin B12 1.4 μg / ml, folic acid 1 μg / ml, L-aspartic acid 5 μg / ml, glutathione 1.2 μg / ml.

[0025] According to the second aspect of the embodiments of the present invention, the present invention provides a preparation method of the cell preservation solution for intravenous infusion as described above. Dissolve sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione in injection water and mix well to obtain the cell preservation solution for intravenous infusion.

[0026] According to the third aspect of the embodiments of the present invention, the present invention provides the application of the cell preservation solution for intravenous infusion as described above in the preparation of intravenous infusion cell drugs.

[0027] Further, the cells are mesenchymal stem cells and hematopoietic stem cells.

[0028] Further, use the cell preservation solution to prepare a cell suspension.

[0029] Further, the number of cells contained in each milliliter of the cell preservation solution is (1 - 2)×10 7 cells.

[0030] Further, the working temperature of the cell preservation solution is 12 - 18°C.

[0031] The embodiments of the present invention have the following advantages:

[0032] (1) High safety: This product is a medical grade injection preparation and is safe to use.

[0033] (2) The number of cells contained in a unit volume of cell protection fluid is large and the concentration is high, which solves the problem of small number of cells in intravenous transfusion of cell drugs.

[0034] (3) Easy to use, can be directly injected into the vein.

[0035] (4) Cryogenic tubes are portable, easy to transport, and simple to operate. They do not require special treatment and can be used directly, saving costs such as site construction and instrument purchase, and are also more convenient for promotion and use.

[0036] (5) The long storage time of cells buys valuable time for the drug application company to test fresh preparations for drug use. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the implementation methods of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the implementation methods or the description of the prior art. Obviously, the drawings in the following description are only exemplary, and for ordinary technicians in this field, other implementation drawings can be derived from the provided drawings without creative work.

[0038] Figure 1 This is a CCK-8 cell proliferation activity detection diagram provided by the present invention;

[0039] Figure 2 A comparison chart of three differentiation tests provided by the present invention;

[0040] Figure 3 This is a flow cytometry image after normal culture provided by the present invention;

[0041] Figure 4 This is a flow cytometry image of cells after being protected and cultured with the protective solution provided by the present invention;

[0042] Figure 5 A cell diagram after normal culture provided by the present invention;

[0043] Figure 6 This is a diagram of cells after being cultured and protected by the cell protection solution provided by the present invention. DETAILED DESCRIPTION

[0044] The following specific embodiments illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0045] For those technical or conditions not specified in the embodiments, they shall be in accordance with the technologies or conditions described in the literature in this field or in accordance with the product specifications. For the reagents or instruments without indicating the manufacturer, they are all conventional products that can be obtained through regular channels.

[0046] 20% Human Albumin: Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd., National Medicine Approval No. S10940028.

[0047] Example 1

[0048] This example provides a cell protection solution for intravenous infusion. By adding sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione into injection water and mixing them evenly.

[0049] Among them, the final concentrations of sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione are successively: 6mg / ml, 0.3mg / ml, 3.1mg / ml, 0.2mg / ml, 0.23μg / ml, 0.02ml / ml, 6μg / ml, 1.4μg / ml, 1μg / ml, 5μg / ml, 1.2μg / ml.

[0050] Example 2

[0051] This example provides a cell protection solution for intravenous infusion. By adding sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione into injection water and mixing them evenly.

[0052] Among them, the final concentrations of sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione are successively: 7.6mg / ml, 0.4mg / ml, 3.4mg / ml, 0.1mg / ml, 0.28μg / ml, 0.03ml / ml, 5μg / ml, 1.8μg / ml, 0.5μg / ml, 7μg / ml, 1.8μg / ml.

[0053] Example 3

[0054] This example provides a cell protective solution for intravenous infusion. Sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione are added to injection water and mixed evenly.

[0055] Among them, the final concentrations of sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione are as follows: 4.5 mg / ml, 0.2 mg / ml, 2.8 mg / ml, 0.3 mg / ml, 0.16 μg / ml, 0.02 ml / ml, 7 μg / ml, 1 μg / ml, 0.8 μg / ml, 4 μg / ml, 0.5 μg / ml.

[0056] Comparative Example 1

[0057] This comparative example provides a cell protective solution for intravenous infusion. The difference from Example 1 is that L-aspartic acid is replaced with an equal amount of glutathione.

[0058] Comparative Example 2

[0059] This comparative example provides a cell protective solution for intravenous infusion. The difference from Example 1 is that the final concentrations of sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid, and glutathione are as follows: 6 mg / ml, 0.3 mg / ml, 2.0 mg / ml, 0.2 mg / ml, 0.23 μg / ml, 0.02 ml / ml, 1 μg / ml, 6 μg / ml, 1 μg / ml, 5 μg / ml, 3 μg / ml.

[0060] Example 4

[0061] This example provides an application of a cell protective solution in the preparation of intravenous infusion cell drugs

[0062] 1. Preparation of bone marrow mesenchymal stem cells

[0063] 1.1 Take 45 ml of bone marrow blood donated by a volunteer, add it to a centrifuge tube and centrifuge at 3000 revolutions for 30 minutes. Use a 2 ml pipette to aspirate the buffy coat layer, dilute it with PBS at a ratio of 1:1 (volume ratio), and add it to a 50 ml centrifuge tube containing the same volume of lymphocyte separation solution. The operation must be slow and the liquid level should not be broken. After paving the bottom, it can be accelerated evenly until the scale reaches 45 ml.

[0064] 1.2 After the separation liquid is completely added, gently place it in a centrifuge and centrifuge at 400 g for 30 min (pay attention to adjusting the deceleration and acceleration). After centrifugation, extract the white layer separated by gradient density centrifugation. Slowly suck it out with a straight tube (the white layer), then make up to 45 ml with PBS and put it into the centrifuge, and centrifuge at 600 g for 5 min.

[0065] 1.3 After centrifugation, pour out the supernatant and leave the required cells. Take an appropriate amount of red blood cell lysate with a straight tube to disperse the cells to achieve the purpose of lysing red blood cells (control the time at about 3 min). After the lysis time, add PBS to 45 ml and put it into the centrifuge, and centrifuge at 600 g for 5 min. After the centrifugation time is up, pour out the waste liquid, add PBS to 45 ml and put it into the centrifuge, and centrifuge at 600 g for 5 minutes.

[0066] 1.4 After centrifugation, disperse the cells with complete medium (α - mem), count them and add them to a 150 cm 2 culture flask. After adding 25 ml of complete medium to each flask, place it in a constant temperature incubator with 5% carbon dioxide and 37 °C.

[0067] 1.5 After static culture for 24 h, pour out the supernatant, then wash the bottom of the flask with 0.9% physiological saline. After washing, add complete medium (α - mem) for liquid change. Observe the cell growth status every day after liquid change, and give liquid change every 3 d until the cells grow to about 80% in the flask. Generally, it can be passaged and amplified to P1 generation in about 10 days. In the later stage, passage once every 5 days until the cell confluence reaches more than 80% at the eighth generation.

[0068] 1.6 After photographing and retaining the cells, perform digestion treatment. Discard the medium in the culture flask, add an appropriate amount of physiological saline to wash the cell surface. After washing, pour out the washing liquid. After sucking out the residual liquid with a 10 ml pipette, add 3 ml of trypsin and digest at room temperature for 3 - 5 min, then use an appropriate amount of termination solution to terminate digestion. Transfer the cell suspension to a 50 ml centrifuge tube, and use physiological saline to rinse the culture flask to collect the residual cells. After washing, transfer them to the 50 ml centrifuge tube in the same way, centrifuge at 500 g for 5 min, and collect the cells.

[0069] 2. Prepare cell suspension

[0070] After centrifugation, rinse the cells with cell preservation solution, centrifuge at 500 g for 5 min and wash twice, then resuspend, filter through a 100 μm mesh sieve, take a sample for counting, and adjust the cell density to (1 - 2)×10 7 / ml to make a cell suspension.

[0071] 3. Preservation

[0072] Place 1 ml of cell suspension into a 5-ml cryopreservation tube with a breathable membrane, and store or transport it using a biological vaccine refrigerator at a temperature of 16°C to obtain a directly usable intravenous infusion cell drug.

[0073] Test Example 1

[0074] Perform re-culture experiments, CCK-8 cell proliferation viability detection, three-lineage differentiation detection, flow cytometry detection, viability detection, quantity detection, etc. on the cells protected by the cell protection solution of the embodiment of the present invention.

[0075] 1. Re-culture experiment

[0076] The cell suspension of Example 4 stored in the cell protection solution of Example 1 for 24 hours was re-attached and cultured with the cells cultured in normal medium. When the cells reached 80% confluence, CCK-8 cell proliferation viability detection and three-lineage differentiation detection were performed.

[0077] The results are shown in Figure 1-2 , and the results show that there is no significant difference in the proliferation viability and differentiation ability of the cells stored in the cell protection solution of the embodiment of the present invention and the cells cultured in normal medium.

[0078] 2. Flow cytometry detection

[0079] The cell suspension of Example 4 stored in the cell protection solution of Example 1 for 24 hours and the cells cultured in normal medium were respectively subcultured until the seventh generation, and flow cytometry was performed using a conventional method. The flow cytometry chart is shown in Figure 3-4 , and the flow cytometry data are shown in Table 1 below, and the cell chart is shown in Figure 5-6 .

[0080] Table 1

[0081]

[0082] The results show that there is no significant difference in the expression of specific markers of the cells stored in the cell protection solution of the embodiment of the present invention and the cells after normal culture.

[0083] 3. Cell quantity and viability detection

[0084] The cell suspension of Example 4 stored in the cell protection solution of Example 1 was tested for cell quantity and viability, and the results of different cell suspensions are shown in Tables 2-3 below.

[0085] Table 2

[0086] Storage time 0h 6h 12h 18h 24h 26h Quantity <![CDATA[1×10 7 / ml]]> <![CDATA[1.05×10 7 / ml]]> <![CDATA[1.08×10 7 / ml]]> <![CDATA[1.06×10 7 / ml]]> <![CDATA[1.02×10 7 / ml]]> <![CDATA[1.08×10 7 / ml]]> Survival rate 98.63％ 97.91％ 98.15％ 97.97％ 97.91％ 98.12％

[0087] Table 3

[0088] Storage time 0h 6h 12h 18h 24h 26h Quantity <![CDATA[2×10 7 / ml]]> <![CDATA[2.03×10 7 / ml]]> <![CDATA[1.97×10 7 / ml]]> <![CDATA[2.01×10 7 / ml]]> <![CDATA[2.05×10 7 / ml]]> <![CDATA[2.07×10 7 / ml]]> Survival rate 98.82％ 98.17％ 97.69％ 97.77％ 98.16％ 97.95％

[0089] The results show that there is no difference in the number and survival rate of cells after being preserved with the cell protective solution of the embodiments of the present invention compared with before preservation.

[0090] Test Example 2

[0091] The cell protective solutions of Examples 1-3 and Comparative Examples 1-2 were formulated into cell suspensions with a cell density of 1.5×10 7 / ml according to the method of Example 4. 1 ml of the cell suspension was placed in a 5-ml cryopreservation tube with a breathable membrane and stored in a biological vaccine refrigerator at 16°C. After 5 days, the cryopreservation tube was taken out, diluted with 5 times the volume of 0.9% normal saline, and the cell survival rate was calculated. The results are shown in Table 4 below.

[0092] Table 4

[0093] Cell protection solution 0d 5d Example 1 98.68％ 98.51％ Example 2 98.25％ 97.48％ Example 3 98.84％ 97.05％ Comparative example 1 98.36％ 91.74％ Comparative example 2 98.52％ 88.57％

[0094] The results show that the cell protective solution of the embodiments of the present invention can maintain the survival rate of cells for a long time and has a better cell protection effect.

[0095] Test Example 3

[0096] 1. Safety detection

[0097] The cell suspension obtained after being preserved with the cell protective solution of Example 1 in Example 4 for 5 days was subjected to safety detections such as sterility, endotoxin, microorganisms, mycoplasma, etc. The results are shown in Table 5 below.

[0098] Table 5

[0099] Property Sample Qualified standard Whether it meets the standard (√ / ×) Residual detection of fetal bovine serum <22 ng / 10^6 cells <50 ng / 10^6 cells √ Sterility test Negative Negative √ Endotoxin test <0.25 EU <0.5 EU √ Mycoplasma test Negative Negative √ Cytomegalovirus test Negative Negative √ Epstein-Barr virus (EBV) test Negative Negative √ Human immunodeficiency virus test Negative Negative √ Hepatitis B virus test Negative Negative √ Hepatitis C virus test Negative Negative √ Syphilis virus test Negative Negative √

[0100] The results show that after the cells are preserved with the cell protective solution of the embodiments of the present invention, no sterility, endotoxin, microorganisms, mycoplasma, etc. are detected, and the safety detection is qualified.

[0101] 2. Animal experiment

[0102] Select 10 batches, a total of 160 (half male and half female) healthy SD rats weighing about 200 g and aged 6 - 9 months. Add them to 0.9% physiological saline according to the dosage to prepare the cell drug. Infuse the cell drug of Example 4 strictly in accordance with the intravenous puncture procedure (using the cell preservation solution of Example 1 with a preservation time of 5 days). Inject 24 (half male and half female) rats in 4 batches with physiological saline as a control. Observe continuously for at least 14 days after injection. The observation indicators include clinical symptoms (such as animal appearance, behavior, diet, response to stimuli, secretions, excretions), death situation (death time, reaction before death, etc.), change in body weight (before injection and at the end of the observation period), and measure the blood routine, blood biochemical indexes, and organ - body ratio, and study and determine their normal reference value ranges. After the end of the observation period, euthanize the animals for dissection. The results are shown in Table 6 below.

[0103] Table 6

[0104]

[0105] The results show that there are no abnormal clinical symptoms before and after injection, no deaths, the body weights of all rats increase, the blood routine, blood biochemical indexes, and organs are all within the normal range, and the volume, color, and texture of the organs are normal without any abnormalities after dissection.

[0106] Although the present invention has been described in detail with general descriptions and specific examples above, based on the present invention, some modifications or improvements can be made, which are obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection required by the present invention.

Claims

1. A cell protection solution for intravenous reinfusion, characterized in that: The cell protection solution includes: sodium chloride 4-8 mg / ml, potassium chloride 0.1-0.5 mg / ml, sodium lactate 2.8-3.5 mg / ml, calcium chloride 0.1-0.3 mg / ml, glucose 0.15-0.3 μg / ml, 20% human albumin 0.01-0.03 ml / ml, vitamin E 4-8 μg / ml, vitamin B12 0.2-2 μg / ml, folic acid 0.5-2 μg / ml, L-aspartic acid 4-8 μg / ml, and glutathione 0.5-2 μg / ml.

2. The cell protection solution for intravenous reinfusion according to claim 1, characterized in that: The cell protection solution includes: sodium chloride 6 mg / ml, potassium chloride 0.3 mg / ml, sodium lactate 3.1 mg / ml, calcium chloride 0.2 mg / ml, glucose 0.23 μg / ml, 20% human albumin 0.02 ml / ml, vitamin E 6 μg / ml, vitamin B12 1.4 μg / ml, folic acid 1 μg / ml, L-aspartic acid 5 μg / ml, and glutathione 1.2 μg / ml.

3. The method for preparing the cell protection solution for intravenous reinfusion according to claim 1 or 2, characterized in that: Sodium chloride, potassium chloride, sodium lactate, calcium chloride, glucose, 20% human albumin, vitamin E, vitamin B12, folic acid, L-aspartic acid and glutathione are dissolved in injection water and mixed to obtain the cell protection solution for intravenous reinfusion.

4. Use of the cell protection solution for intravenous reinfusion according to claim 1 or 2 in the preparation of intravenous reinfusion cell medicine.

5. The use according to claim 4, characterized in that: The cells are mesenchymal stem cells and hematopoietic stem cells.

6. The use according to claim 4, characterized in that: The cells are prepared into a cell suspension using the cell protection solution.

7. The use according to claim 6, characterized in that: The number of cells contained in each ml of the cell protection solution is (1-2)×10 7 indivual.

8. The use according to claim 6, characterized in that: The working temperature of the cell protection solution is 12-18°C.

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