Tumor vaccine based on DC cells and preparation method thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses a DC cell-based tumor vaccine and a preparation method thereof, and the preparation method comprises the following steps: (1) separating peripheral blood to prepare a mononuclear cell suspension; (2) inoculating the mononuclear cell suspension into a first culture medium, culturing for 48-72 hours, then replacing 40-60% of the first culture medium every 48-72 hours, and continuing culturing to obtain a first cell sap; (3) preparing the tumor tissue into a cell lysate; and (4) adding the cell lysate, polyinosinic-cytidylic acid, TNF-alpha and alpha-ketoglutaric acid into the first cell sap, and carrying out co-culture to obtain the DC cell-based tumor vaccine. The DC cell-based tumor vaccine prepared by the method provided by the invention can specifically kill tumor cells.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a DC cell-based tumor vaccine and a preparation method thereof. Background Art

[0002] Tumor immunotherapy has become the fourth largest tumor treatment method after surgery, radiotherapy, and chemotherapy. Traditional tumor treatment methods have problems such as poor targeting and easy recurrence. DC vaccines can specifically identify and kill tumor cells by activating the patient's own immune system, and have better tumor targeting, safety, and low toxicity.

[0003] Dendritic cells (DCs) are important immune cells with powerful antigen-presenting abilities, playing a key role in activating initial T cell responses. In recent years, DC-based tumor vaccines have been recognized as a promising avenue for tumor immunotherapy. DCs can ingest tumor antigens, process them, and present them to T cells, thereby initiating specific anti-tumor immune responses.

[0004] In view of this, this application is filed. Summary of the Invention

[0005] The present invention provides a DC cell-based tumor vaccine and a preparation method thereof. The DC cell-based tumor vaccine prepared by the method of the present invention can specifically kill tumor cells.

[0006] The present invention solves its technical problems by adopting the following technical solutions: A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) Isolate and prepare mononuclear cell suspension from peripheral blood; (2) Inoculating the mononuclear cell suspension into the first culture medium and culturing for 48 to 72 hours, then replacing 40 to 60% of the first culture medium every 48 to 72 hours and continuing the culture to obtain the first cell fluid; (3) preparing tumor tissue into cell lysates; (4) Adding cell lysate, polyinosinic-polycytidylic acid, TNF-α, and α-ketoglutaric acid to the first cell solution for co-culture to obtain a DC cell-based tumor vaccine.

[0007] As a preferred embodiment of the present invention, the culture is carried out in an incubator at 36.5-37.5°C and 5% CO2.

[0008] As a preferred embodiment of the present invention, the cell concentration in the mononuclear cell suspension is 1-2×10 6 pieces / mL.

[0009] As a preferred embodiment of the present invention, the first culture medium comprises the following components: 4-10 mg / mL isoquercetin, 4-10 mg / mL glutamine, 100-300 μg / mL IL-7, 100-300 μg / mL IL-4, 2-10 mM N-acetylcysteine, 10-40 μM β-mercaptoethanol, 100-200 ng / mL linolenic acid, and XVIVO-15 serum-free medium.

[0010] As a preferred embodiment of the present invention, the culturing time in step (2) is 8 to 12 days.

[0011] As a preferred embodiment of the present invention, the method for preparing tumor tissue into cell lysate is: S1. Take the tumor tissue, mince it, add digestive enzymes to digest it, resuspend it in RPMI1640 medium, centrifuge it, remove the supernatant, and resuspend it in RPMI1640 medium. Adjust the cell density to 0.5~2×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 10-30 seconds and thaw in a 36-38°C water bath for 8-12 minutes. S3. Repeat step S2 4 to 6 times to obtain cell lysate.

[0012] As a preferred embodiment of the present invention, the culturing time in step (4) is 8 to 20 hours.

[0013] As a preferred embodiment of the present invention, the volume ratio of the cell lysate to the first cell fluid is (0.4-0.8):1.

[0014] As a preferred embodiment of the present invention, the final concentration of the polyinosinic-polycytidylic acid is 4-6 mg / mL; The final concentration of the α-ketoglutaric acid is 1-3 mg / mL.

[0015] As a preferred embodiment of the present invention, the final concentration of TNF-α is 200-500 μg / mL.

[0016] The present invention also provides a DC cell-based tumor vaccine, which is prepared using the above-mentioned preparation method.

[0017] The beneficial effects of the present invention are as follows: (1) The present invention utilizes the patient's own tumor tissue sample to prepare tumor antigens, thereby achieving personalized customization of DC vaccines and improving the pertinence and effectiveness of treatment. (2) The present invention optimizes the first culture medium and the added components of the co-culture to provide sufficient nutritional support for DC cells, promote their growth and proliferation, promote the complete maturation of DC cells, improve the DC cells' ability to absorb and process tumor antigens, increase the antigen loading capacity and immune activation effect, enhance the immune activation effect, and at the same time enhance the antigen presentation ability of DC cells. By combining with negatively charged antigens and DC cell membranes, the endocytosis and presentation of antigens are promoted, effectively improving the tumor cell killing rate. DETAILED DESCRIPTION

[0018] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0019] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0020] In the present invention, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Furthermore, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0021] In the present invention, there is no particular limitation on the specific dispersion and stirring treatment methods.

[0022] The reagents or instruments used in the present invention without indicating the manufacturer are all conventional products that can be obtained commercially. The raw materials used in the comparative examples and the raw materials used in the parallel experiments of the examples are the same commercially available products unless otherwise specified.

[0023] Example 1 A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6 / mL, and a mononuclear cell suspension was obtained; (2) The mononuclear cell suspension was inoculated into the first culture medium and cultured for 2 days, and then 50% of the first culture medium was replaced every 2 days for another 8 days to obtain the first cell fluid; The first culture medium comprises the following components: 8 mg / mL isoquercetin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetylcysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the remainder XVIVO-15 serum-free culture medium.

[0024] (3) preparing cell lysates from lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0025] (4) Adding the cell lysate, polyinosinic-cytidylic acid (PI), TNF-α, and α-ketoglutaric acid to the first cell solution and co-culturing for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of the PI is 5 mg / mL; the final concentration of the α-ketoglutaric acid is 2 mg / mL; and the final concentration of the TNF-α is 400 μg / mL.

[0026] Example 2 A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6 / mL, and a mononuclear cell suspension was obtained; (2) The mononuclear cell suspension was inoculated into the first culture medium and cultured for 2 days, and then 50% of the first culture medium was replaced every 2 days for another 8 days to obtain the first cell fluid; The first culture medium comprises the following components: 8 mg / mL isoquercitrin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetyl cysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the balance of XVIVO-15 serum-free medium.

[0027] (3) preparing a cell lysate from the lung cancer tumor tissue; S1, taking lung cancer tumor tissue, removing necrotic tissue and non-tumor tissue adjacent to the cancer, cutting, adding digestive enzymes for digestion, resuspending with RPMI1640 medium, centrifuging, removing the supernatant, resuspending with RPMI1640 medium again, adjusting the cell density to 1×10 7 cells / ml, to obtain a cell solution; S2, freezing the cell solution with liquid nitrogen for 25 s, and thawing at 37°C water bath for 10 min; S3, repeating the step S2 for 5 times to obtain a cell lysate.

[0028] (4) adding the cell lysate, poly I:C, TNF-α, and α-ketoglutaric acid to the first cell solution for co-culture for 12 h to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.4:1. The final concentration of the poly I:C is 4 mg / mL; the final concentration of the α-ketoglutaric acid is 3 mg / mL; and the final concentration of the TNF-α is 200 μg / mL.

[0029] Example 3 A preparation method of a DC cell-based tumor vaccine, comprising the following steps: (1) collecting 20 mL of peripheral blood of a lung cancer patient, separating mononuclear cells by Ficoll density gradient centrifugation, and adjusting the density of the mononuclear cells to 1×10 6 cells / mL with lymphocyte culture solution GT-T551 (also referred to as GT-T551 culture solution) to obtain a mononuclear cell suspension; (2) inoculating the mononuclear cell suspension into a first culture medium for culture for 2 days, then replacing 50% of the first culture medium every 2 days for continued culture for 8 days to obtain a first cell solution; The first culture medium comprises the following components: 8 mg / mL isoquercitrin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetyl cysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the balance of XVIVO-15 serum-free medium.

[0030] (3) preparing a cell lysate from the lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0031] (4) Adding the cell lysate, polyinosinic-cytidylic acid (PI), TNF-α, and α-ketoglutaric acid to the first cell solution and co-culturing for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.6:1. The final concentration of the PI is 6 mg / mL; the final concentration of the α-ketoglutaric acid is 1 mg / mL; and the final concentration of the TNF-α is 500 μg / mL.

[0032] Comparative Example 1

[0033] A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6 / mL, and a mononuclear cell suspension was obtained; (2) The mononuclear cell suspension was inoculated into the first culture medium and cultured for 2 days, and then 50% of the first culture medium was replaced every 2 days for another 8 days to obtain the first cell fluid; The first culture medium includes the following components: 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetylcysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the remainder XVIVO-15 serum-free culture medium.

[0034] (3) preparing cell lysates from lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0035] (4) Adding the cell lysate, polyinosinic-cytidylic acid (PI), TNF-α, and α-ketoglutaric acid to the first cell solution and co-culturing for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of the PI is 5 mg / mL; the final concentration of the α-ketoglutaric acid is 2 mg / mL; and the final concentration of the TNF-α is 400 μg / mL.

[0036] Comparative Example 2

[0037] A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6 / mL, and a mononuclear cell suspension was obtained; (2) The mononuclear cell suspension was inoculated into the first culture medium and cultured for 2 days, and then 50% of the first culture medium was replaced every 2 days for another 8 days to obtain the first cell fluid; The first culture medium comprises the following components: 8 mg / mL isoquercetin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetylcysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the remainder XVIVO-15 serum-free culture medium.

[0038] (3) preparing cell lysates from lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0039] (4) Adding the cell lysate, TNF-α, and α-ketoglutarate to the first cell solution and co-culturing for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of the α-ketoglutarate is 2 mg / mL; the final concentration of the TNF-α is 400 μg / mL.

[0040] Comparative Example 3

[0041] A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6 / mL, and a mononuclear cell suspension was obtained; (2) The mononuclear cell suspension was inoculated into the first culture medium and cultured for 2 days, and then 50% of the first culture medium was replaced every 2 days for another 8 days to obtain the first cell fluid; The first culture medium comprises the following components: 8 mg / mL isoquercetin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetylcysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the remainder XVIVO-15 serum-free culture medium.

[0042] (3) preparing cell lysates from lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0043] (4) Adding the cell lysate, polyinosinic-polycytidylic acid (PIC), and TNF-α to the first cell solution for co-culture for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of the PIP is 5 mg / mL; the final concentration of the TNF-α is 400 μg / mL.

[0044] Comparative Example 4

[0045] A method for preparing a tumor vaccine based on DC cells comprises the following steps: (1) 20 mL of peripheral blood was collected from lung cancer patients, and mononuclear cells were isolated by Ficoll density gradient centrifugation. The density of mononuclear cells was adjusted to 1×10 using lymphocyte culture medium GT-T551 (also known as GT-T551 culture medium). 6cells / mL, to obtain a single-nucleus cell suspension; (2) inoculate the single-nucleus cell suspension into the first culture medium and culture for 2 days, then replace 50% of the first culture medium every 2 days and continue to culture for 8 days, to obtain a first cell solution; The first culture medium comprises the following components: 8 mg / mL isoquercitrin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetyl cysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the balance of XVIVO-15 serum-free medium.

[0046] (3) prepare a cell lysate from the lung cancer tumor tissue; S1, take the lung cancer tumor tissue, remove the necrotic tissue and non-tumor tissue adjacent to the cancer, cut into small pieces, add digestive enzymes for digestion, resuspend with RPMI1640 medium, centrifuge, remove the supernatant, resuspend with RPMI1640 medium, adjust the cell density to 1×10 7 cells / mL, to obtain a cell solution; S2, freeze the cell solution in liquid nitrogen for 25 s, and thaw at 37°C water bath for 10 min; S3, repeat S2 for 5 times, to obtain a cell lysate.

[0047] (4) add the cell lysate, poly I:C, and TNF-α to the first cell solution for co-culture for 12 h, to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of poly I:C is 7 mg / mL; and the final concentration of TNF-α is 400 μg / mL.

[0048] Comparative Example 5

[0049] A preparation method of a DC cell-based tumor vaccine, comprising the following steps: (1) collect 20 mL of peripheral blood from a lung cancer patient, separate mononuclear cells by Ficoll density gradient centrifugation, and adjust the density of the single-nucleus cells to 1×10 6 cells / mL, to obtain a single-nucleus cell suspension; (2) inoculate the single-nucleus cell suspension into the first culture medium and culture for 2 days, then replace 50% of the first culture medium every 2 days and continue to culture for 8 days, to obtain a first cell solution; The first culture medium comprises the following components: 8 mg / mL isoquercetin, 5 mg / mL glutamine, 200 μg / mL IL-7, 200 μg / mL IL-4, 5 mM N-acetylcysteine, 30 μM β-mercaptoethanol, 120 ng / mL linolenic acid, and the remainder XVIVO-15 serum-free culture medium.

[0050] (3) preparing cell lysates from lung cancer tumor tissue; S1. Take lung cancer tumor tissue, remove necrotic tissue and adjacent non-tumor tissue, mince, add digestive enzymes for digestion, resuspend in RPMI1640 medium, centrifuge, remove supernatant, and resuspend in RPMI1640 medium. Adjust the cell density to 1×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 25 seconds and thaw in a 37°C water bath for 10 minutes. S3. Repeat step S2 5 times to obtain cell lysate.

[0051] (4) Adding the cell lysate, TNF-α, and α-ketoglutarate to the first cell solution and co-culturing for 12 hours to obtain a DC cell-based tumor vaccine. The volume ratio of the cell lysate to the first cell solution is 0.5:1. The final concentration of the α-ketoglutarate is 7 mg / mL; the final concentration of the TNF-α is 400 μg / mL.

[0052] Test Case The tumor vaccine prepared above was co-cultured with primary tumor cells at a ratio of 5:1, and the tumor killing rate (%) was calculated after 24 hours of culture.

[0053] Table 1

[0054] As can be seen from Table 1, the DC cell-based tumor vaccine prepared by the method of the present invention can specifically kill tumor cells.

[0055] Finally, it should be noted that the above embodiments are intended to illustrate the technical solutions of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for preparing a tumor vaccine based on DC cells, characterized in that: The following steps are involved: (1) Isolate and prepare mononuclear cell suspension from peripheral blood; (2) Inoculating the mononuclear cell suspension into the first culture medium and culturing for 48 to 72 hours, then replacing 40 to 60% of the first culture medium every 48 to 72 hours and continuing the culture to obtain the first cell fluid; (3) preparing tumor tissue into cell lysates; (4) Adding cell lysate, polyinosinic-polycytidylic acid, TNF-α, and α-ketoglutaric acid to the first cell solution for co-culture to obtain a DC cell-based tumor vaccine.

2. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The culture was carried out in an incubator at 36.5-37.5°C and 5% CO2.

3. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The cell concentration in the mononuclear cell suspension is 1-2×10 6 pieces / mL.

4. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The first culture medium comprises the following components: 4-10 mg / mL isoquercetin, 4-10 mg / mL glutamine, 100-300 μg / mL IL-7, 100-300 μg / mL IL-4, 2-10 mM N-acetylcysteine, 10-40 μM β-mercaptoethanol, 100-200 ng / mL linolenic acid, and XVIVO-15 serum-free culture medium.

5. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The culture time in step (2) is 8 to 12 days.

6. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The method for preparing tumor tissue into cell lysate is: S1. Take the tumor tissue, mince it, add digestive enzymes to digest it, resuspend it in RPMI1640 medium, centrifuge it, remove the supernatant, and resuspend it in RPMI1640 medium. Adjust the cell density to 0.5~2×10 7 / ml, and cell fluid was obtained; S2. Freeze the cell suspension with liquid nitrogen for 10-30 seconds and thaw in a 36-38°C water bath for 8-12 minutes. S3. Repeat step S2 4 to 6 times to obtain cell lysate.

7. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The culture time in step (4) is 8 to 20 hours.

8. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The volume ratio of the cell lysate to the first cell solution is (0.4-0.8):

1.

9. The method for preparing a tumor vaccine based on DC cells according to claim 1, characterized in that: The final concentration of the polyinosinic-polycytidylic acid is 4-6 mg / mL; The final concentration of the α-ketoglutaric acid is 1-3 mg / mL; The final concentration of TNF-α is 200-500 μg / mL.

10. A tumor vaccine based on DC cells, characterized in that: The invention is prepared by the preparation method according to any one of claims 1 to 9.

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