Dendritic cells that can target cancer stem cells and their combination with t cells, and preparation method and use thereof
By developing dendritic cells that target cancer stem cells and combining them with T cells, the method effectively addresses the ineffectiveness of existing immunotherapy in eliminating cancer stem cells, thereby improving cancer treatment outcomes.
Patent Information
- Application Number
- PCT/IB2024/000649
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2023-11-15
- Filing Date
- 2024-12-12
- Publication Date
- 2025-05-22
AI Technical Summary
Existing immunotherapy methods are ineffective in targeting and eliminating cancer stem cells, leading to drug resistance, recurrence, and metastasis in tumors.
Development of dendritic cells that can target cancer stem cells, combined with T cells, through isolation, culture, and antigen presentation to specifically recognize and kill cancer stem cells.
The combination of dendritic cells and T cells effectively eliminates cancer stem cells, preventing tumor drug resistance, recurrence, and metastasis, thereby enhancing cancer treatment efficacy.
Smart Images
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Abstract
Description
[0001] Dendritic Cells that can Target Cancer Stem Cells and their Combination with T Cells, and Preparation Method and Use thereof
[0002] BACKGROUND OF THE INVENTION
[0003] 1. FIELD OF THE PRESENT DISCLOSURE
[0004] The present disclosure relates to a dendritic cell and its combination with T cells, as well as its preparation method and use thereof, particularly to a dendritic cell that can target cancer stem cells and its combination with T cells, as well as its preparation method and use thereof.
[0005] 2. BACK GROUND
[0006] Immunotherapy is a major breakthrough in the field of cancer treatment in recent years, which involves activating and enhancing the body’s own immune system to combat cancer. Compared with traditional chemotherapy and radiation therapy, immunotherapy has the advantages of fewer side effects and long-lasting efficacy.
[0007] The current immunotherapy mainly includes the following categories:
[0008] 1. Immune checkpoint inhibitors: these drugs activate T cells to attack tumors by blocking the immune suppression pathways utilized by cancer cells, such as PD-l / PD- L1 and CTLA-4. Its representative drugs include Opdivo® (also known as nivolumab) and Keytruda® (also known as pembrolizumab).
[0009] 2. CAR-T cell therapy: the patient’s own T cells are genetically engineered to specifically recognize and kill cancer cells.
[0010] 3. Cancer vaccine: inducing the human body to produce an immune response against tumors by injecting tumor antigens or genetically engineered viruses.
[0011] 4. Cytokine therapy: using cytokines such as IL-2 to enhance the activity of immune cells. 5. Monoclonal antibodies: using drugs such as Rituximab, they can specifically bind to tumor cell surface antigens and activate the immune system.
[0012] Although existing immunotherapy has shown excellent efficacy in certain types of cancer, there are still some obvious limitations, such as the inability to effectively kill drug-resistant, recurrent, or metastatic cancer cells. The crux of the problem lies in the fact that existing immunotherapy mainly targets cancer cells rather than cancer stem cells. As long as cancer stem cells still exist, they will make cancer cells resistant and repeat proliferation or metastasis, making the tumor unable to be effectively treated.
[0013] Therefore, it is necessary to provide a T cell that can specifically recognize and kill cancer stem cells to solve the problem of ineffective treatment of tumors due to drug resistance, recurrence, or metastasis.
[0014] SUMMARY OF THE INVENTION
[0015] In view of the above problems, the inventor of the present disclosure has considered and invented a dendritic cell that can target cancer stem cells, a combination thereof with T cells, a preparation method and use thereof, to improve the problem of ineffective treatment of cancer.
[0016] The first objective of the present disclosure is to provide dendritic cells that can target cancer stem cells and their preparation method. The dendritic cells that can target cancer stem cells are first isolated from peripheral blood collected from the human body and induced to culture into dendritic cells (DCs). Then, cancer stem cell surface antigens are extracted from cancer stem cells (CSCs) and co-cultured with the dendritic cells to obtain dendritic cells that can target cancer stem cells.
[0017] The second objective of the present disclosure is to provide a combination of dendritic cells and T cells that can target cancer stem cells, and a preparation method thereof. The combination of dendritic cells and T cells that can target cancer stem cells is obtained by first isolating T cells from peripheral blood collected from the human body and co culturing them with dendritic cells that can target cancer stem cells as described above, thereby obtaining the combination of dendritic cells and T cells that can target cancer stem cells.
[0018] The third objective of the present disclosure is to provide a combination of dendritic cells and T cells that can target cancer stem cells, which is used to eliminate cancer stem cells and prevent tumors from developing drug resistance, recurrence, and metastasis. Inject the dendritic cells and T cells that can target cancer stem cells back into cancer patients, and effectively eliminate cancer stem cells by utilizing their surface antigen properties that can target cancer stem cells.
[0019] Below, specific embodiments will be provided in conjunction with the accompanying drawings to provide a detailed explanation of the purpose, technical content, features, and achieved effects of the present disclosure.
[0020] BRIEF DESCRIPTION OF THE DRAWINGS
[0021] To describe the technical solutions in the embodiments of this application more clearly, the following briefly introduces the accompanying drawings required for describing the embodiments. Apparently, the accompanying drawings in the following description show merely some embodiments of this application, and a person of ordinary skill in the art may still derive other drawings from these accompanying drawings without creative efforts.
[0022] Fig. 1 is a schematic diagram of a preparation process of dendritic cells that can target cancer stem cells and their combination with T cells according to one embodiment of the present disclosure.
[0023] Fig. 2 is a schematic diagram of the content of CD14+monocytes before induction into CDllc+dendritic cells according to one embodiment of the present disclosure. Fig. 3 is a schematic diagram of the content of CD14+monocytes and CDl lc+dendritic cells after induction of CD14+monocytes into CDl lc+dendritic cells according to one embodiment of the present disclosure.
[0024] Fig. 4 is a schematic diagram of CD14+monocytes forming CD1 lc+dendritic cells before and after induction, observed under a microscope according to one embodiment of the present disclosure.
[0025] Fig. 5 is a schematic diagram of the content of CD8+T cells before and after coculturing with dendritic cells that can target cancer stem cells according to one embodiment of the present disclosure.
[0026] Fig. 6 is a schematic diagram of the toxic effect on the tumorigenic cancer stem cells of human pancreatic cancer by using the vesicles, antigens or exosomes of the parental tumor cancer cells of human pancreatic cancer cells or their tumorigenic cancer stem cells as antigen presentation, co-culturing with dendritic cells to obtain dendritic cells that can target cancer cells or cancer stem cells, and then co-culturing dendritic cells of target cancer cells or cancer stem cells with T cells to obtain the combination of target cancer cells or dendritic cells of cancer stem cells and T cells according to one embodiment of the present disclosure.
[0027] Fig. 7 is a diagram of the toxic effect of the increased toxic T cells targeting WT1 antigen on human pancreatic cancer cells and tumorigenic cancer stem cells after the dendritic cells that can target cancer stem cells and their combinations with T cells, and the dendritic cells co cultured with WT1 antigen acquire dendritic cells that can target WT1 antigen ability, and then inject dendritic cells that can target WT1 antigen ability into the body of cancer patients, according to one embodiment of the present disclosure.
[0028] DESCRIPTION OF THE EMBODIMENTS
[0029] In order to facilitate the understanding of the technical features, content, advantages, and achievable effects of the present disclosure by the examiner, the present disclosure is hereby described in detail in the form of embodiments in conjunction with the accompanying drawings. The drawings used herein are for illustrative and auxiliary purposes only, and may not be the true proportion and precise configuration of the present disclosure after implementation. Therefore, the proportion and configuration relationship of the attached drawings should not be interpreted or limited to the scope of the rights of the present disclosure in practical implementation, and should not be interpreted or limited.
[0030] Unless otherwise defined, all terms used in this article (including technical and scientific terms) have meanings that are commonly understood by those skilled in the art to which the present disclosure belongs. It will be further understood that terms such as those defined in commonly used dictionaries should be interpreted as having meanings consistent with their meanings in the context of the relevant technology and the present disclosure, and will not be interpreted as idealized or overly formal unless explicitly defined in this context.
[0031] Preparation Example 1
[0032] Firstly, please refer to Fig. 1, which is a schematic diagram of a preparation process of dendritic cells that can target cancer stem cells and their combination with T cells according to one embodiment of the present disclosure. The steps are described as follows:
[0033] SI : separating a plurality of monocytes and a plurality of T cells from a peripheral blood collected from a human body.
[0034] In the step SI, a plurality peripheral blood mononuclear cells (PBMCs) were collected from the human body using a plastic pipette and centrifuged at 800 g for 15 minutes in a centrifuge tube containing a density gradient medium (Ficoll®-Paque).
[0035] Next, draw the peripheral blood mononuclear cells into a new sterile centrifuge tube, add phosphate buffered saline (PBS) and mix evenly. Centrifuge at 800 g for 8 minutes and repeat three times to thoroughly clean and remove impurities.
[0036] Next, remove the supernatant after centrifugation, mix the cells evenly with culture medium, count to confirm the number of cells, and add an appropriate amount of fresh culture medium to adjust the cell concentration to 2 * 107cells / mL. Then transfer the cells and culture medium to a cell culture bottle.
[0037] Finally, add MACS® anti-CD14, and anti-CD8, which are two types of magnetic beads (microbeads), were added to cell culture bottles and cultured in an incubator to isolate the required CD14+monocytes and CD8+T cells.
[0038] S2: cultivating the monocytes into a plurality of dendritic cells.
[0039] In the step S2, resuspend the CD14+monocytes obtained in step SI in AIM V™ in the culture medium, after adjusting the cell concentration to 1 x 106cells / mL, transfer the CD14+monocytes and the culture medium to the cell culture bottle, and take out some of them for detection with Flow Cytometry to confirm the content ratio of CD14+monocytes. Please refer to Fig. 2, which is a schematic diagram of the content of CD14+monocytes before induction into CDllc+dendritic cells according to one embodiment of the present disclosure. It can be seen from the figure that after staining with fluorescein isothiocyanate (FITC), the overall content of CD14+monocytes is calculated to be approximately 38.6%.
[0040] Next, IL-4 (Interleukin-4, 500 U / mL) and GM-CSF (Granulocyte-macrophage colony-stimulating factor, 500 U / mL) were added to the cell culture bottle and mixed thoroughly. The mixture was then placed in a culture incubator at 37 °C and 5% CO2 for cultivation. On the third or fourth day of cultivation, aspirate the old culture medium from the bottle, replace it with fresh culture medium containing IL-4 and GM-CSF, and continue to culture in the incubator under the same conditions.
[0041] Next, on the 7thor 8thday of cultivation, TNF-a (2.6 ng / mL) was added to promote dendritic cell maturation, and the cell culture bottle was returned to the incubator for further cultivation for 2 to 3 days. On the 9thor 10thday, extract a portion from the bottle and stain the cells with CDl lc antibody. Then, use a flow cytometer to confirm the proportion of CDllc+dendritic cells. Finally, isolate and purify the required CDllc+dendritic cells.
[0042] Please refer to Fig. 3, which is a schematic diagram of the content of CD14+monocytes and CDllc+dendritic cells after induction of CD14+monocytes into CDllc+dendritic cells according to one embodiment of the present disclosure. From Fig. 3(b), it can be seen that the overall content of CD14+monocytes decreased to about 0.5%. Furthermore, from Fig. 3(b), it can be seen that after staining with phycoerythrin (PE), the overall content of CDllc+dendritic cells was calculated to be about 56.4%, indicating that the majority of CD14+monocytes were indeed induced to transform into CDllc+dendritic cells.
[0043] Please also refer to Fig. 4, which is a schematic diagram of CD14+monocytes forming CDllc+dendritic cells before and after induction, observed under a microscope according to one embodiment of the present disclosure. From Fig. 4(a), it can be seen that before induction, CD1 lc+dendritic cells have various shapes, and cells with a shape close to circular are called CD14+monocytes. From Fig. 4(b), it can be seen that after induction, CD14+monocytes formed CDllc+dendritic cells and were screened. It was found that most of the CD14+monocytes did indeed transform into CDllc+dendritic cells.
[0044] S3 : extracting a plurality of cancer stem cell surface antigens from a plurality of cancer stem cells.
[0045] In the step S3, transfer the supernatant above the cultured cancer stem cells to a centrifuge tube, centrifuge at 2,000 g for 30 minutes to remove cell debris, and then transfer the supernatant containing cell-free culture medium to a new centrifuge tube. Next, add about 0.5 times the volume of extracellular vesicle separation reagent relative to the cell-free culture medium, mix well, and place in a refrigerator at 2 to 8 °C overnight for reaction. The next day, remove it and centrifuge it at 10,000 g for 1 hour to collect the precipitated cancer stem cell exosomes. Resuspend them in an appropriate amount of PBS as the surface antigen presentation of cancer stem cells. Alternatively, chemical pressure agents such as paraformaldehyde (PFA) and dithiothreitol (DTT) can be added to the supernatant to induce apoptosis of cancer stem cells, leading to membrane foaming and the production of approximately 1 to 3 pm sized apoptotic bodies, known as giant plasma membrane vesicles (GPMVs). GPMVs can then be extruded and purified to form a single layer of nano plasma membrane vesicles (nPMVs), which can be used as a surface antigen presentation for cancer stem cells.
[0046] Alternatively, the cultured cancer stem cells can be frozen at a low temperature of approximately -15 °C in a refrigerator, melted at room temperature, placed in liquid nitrogen for 7 minutes, and then placed in a water bath at 37 °C for 7 minutes for freezethaw. After repeated freeze-thaw cycles to rupture the cell wall of cancer stem cells, they were centrifuged and their cancer stem cell antigens were collected as the surface antigen presentation of cancer stem cells.
[0047] S4: co-culturing each of the cancer stem cell surface antigens with the dendritic cells to obtain a plurality of dendritic cells that can target the cancer stem cells.
[0048] In the step S4, the CD1 lc+dendritic cells obtained in the step S2 were resuspended in AIM V™ medium, the cell concentration was adjusted to 1 x 106cells / mL, an appropriate amount of cancer stem cell surface antigen presentation obtained in the step S3 was added, and the CD1 lc+dendritic cells were placed in the incubator at 37 °C and 5% CO2 for overnight co culture, so that CDllc+dendritic cells could present cancer stem cell surface antigen. Next, the CDllc+dendritic cells cultured overnight were centrifuged to remove the supernatant, washed three times with phosphate buffered saline (PBS), and resuspended in AIM V™ medium to obtain CDllc+dendritic cells that can target cancer stem cells.
[0049] S5: co-culturing the dendritic cells that can target cancer stem cells with the T cells to obtain a combination of dendritic cells that can target the cancer stem cells with T cells.
[0050] In the step S5, the CD8+T cells obtained in the step SI were resuspended in AIM V™ medium, the cell concentration was adjusted to 1 x 106cells / mL, an appropriate amount of CDllc+dendritic cells obtained in the step S4 that can target cancer stem cells were added, and incubated in the incubator at 37 °C and 5% CO2 for a total of 1 day, so that CD8+T cells can recognize the cancer stem cell surface antigen presented on CDllc+dendritic cells. The CD8+T cells that can recognize the cancer stem cell surface antigen presented on CDllc+dendritic cells are the combination of dendritic cells that can target the cancer stem cells with T cells.
[0051] Preferably, the ratio of CDllc+dendritic cells targeting cancer stem cells to CD8+T cells used in co culture is 1 : 10.
[0052] Preferably, in order to promote the large-scale proliferation of dendritic cells and T cells that can target cancer stem cells, after co culturing for 1 day, the dendritic cells and T cells that can target cancer stem cells are isolated, and an appropriate amount of IL-2 (Interleukin 2, 200 U / mL) is added for cultivation. During this period, the culture medium containing fresh IL-2 is changed every 2 days, and the culture is continued until the required number of cells are reached, completing the large-scale proliferation of dendritic cells and T cells that can target cancer stem cells.
[0053] Please refer to Fig. 5, which is a schematic diagram of the content of CD8+T cells before and after co-culturing with dendritic cells that can target cancer stem cells according to one embodiment of the present disclosure. As can be seen from the figure, under the induction of CDllc+dendritic cells targeting cancer stem cells, the overall content of CD8+T cells increased from about 24.639% to about 57.013%, an increase of about 2.31 times. In addition, CD8+T cells induced by CDllc+dendritic cells targeting cancer stem cells (z.e., the combination of dendritic cells that can target the cancer stem cells with T cells) can recognize the cancer stem cell surface antigens and specifically poison them, thereby solving the problem of ineffective treatment of tumors due to drug resistance, recurrence, or metastasis.
[0054] Experimental Example 1
[0055] In order to confirm whether the combination of dendritic cells that can target the cancer stem cells with T cells requested by the invention has obvious effect on the use of cancer stem cells, and whether the combination of dendritic cells that can target the cancer stem cells with T cells obtained by culturing cancer stem cell surface antigen presenters from different sources (namely cancer stem cell vesicles, cancer stem cell antigens, and cancer stem cell exocrines) has toxic effect, take human pancreatic cancer cell (PANC-1) as an example, we compared the target cancer cells or the combination of dendritic cells that can target the cancer stem cells with T cells derived from parental tumor cancer cells (PT) or their tumorigenic cancer stem cells (TCSC).
[0056] Please refer to Fig. 6, which is a schematic diagram of the cytotoxic effect on the tumorigenic cancer stem cells of human pancreatic cancer by using the vesicles, antigens or exosomes of the parental tumor cancer cells of human pancreatic cancer cells or their tumorigenic cancer stem cells as antigen presentation, co-culturing with dendritic cells to obtain dendritic cells that can target cancer cells or cancer stem cells, and then co-culturing dendritic cells of target cancer cells or cancer stem cells with T cells to obtain the combination of target cancer cells or dendritic cells of cancer stem cells and T cells according to one embodiment of the present disclosure. It can be seen from Fig. 6(a) that the combination of dendritic cells that can target the cancer stem cells with T cells, or untrained dendritic cells (untrained DC, i.e. dendritic cells co cultured without any antigen presentation), which are obtained by culturing cancer stem cells without adding T cells as the control group, whether with the parental tumor cell vesicles, antigens, or PT exosomes as the source of antigen presentation, for the survival rate of human pancreatic cancer oncogenic cancer stem cells has no significant effect, which means that the combination of dendritic cells that can target the cancer stem cells with T cells cultured with general parental tumor cancer cells as antigen presenters is completely unable to cause cytotoxic effect on oncogenic cancer stem cells.
[0057] It can be seen from Fig. 6(b) that the combination of dendritic cells and T cells that can target cancer stem cells, whether cultured with tumor forming cancer stem cells’ vesicles, antigens or exosomes as the source of antigen presentation, has a significant toxic effect on the survival rate of tumor forming cancer stem cells of human pancreatic cancer, especially when cultured with tumorigenic cancer stem cells’ exosomes, taking tumorigenic cancer stem cells (TCSC only) as the control group. The combination of dendritic cells and T cells cultured as antigen presenters, which can target cancer cells, has the best cytotoxic effect on the tumorigenic cancer stem cells of human pancreatic cancer.
[0058] Experimental Example 2
[0059] According to the results of experimental example 1, it can be found that the combination of dendritic cells that can target the cancer stem cells with T cells cultured with tumor forming cancer stem cells also from human pancreatic cancer cells as the source of antigen presentation, compared with the combination of dendritic cells that can target the cancer stem cells with T cells cultured with parental tumor cancer cells as antigenic presentation, the combination of combination of dendritic cells that can target the cancer stem cells with T cells indeed has an obvious effect of poisoning tumorigenic cancer stem cells of human pancreatic cancer.
[0060] Furthermore, it is further confirmed whether the combination of dendritic cells that can target the cancer stem cells with T cells as claimed in the present disclosure, compared to the existing use of WT1 (Wilms’ tumor gene 1) antigen (one of the specific antigens expressed in most cancer cells) as an antigen presentation and coculture with dendritic cells to obtain dendritic cells capable of targeting WT1 antigen and injecting them back into the bodies of cancer patients to increase the killing effect of WT1 antigen on cancer stem cells, also has a significant effect on the killing of cancer stem cells.
[0061] Please refer to Fig. 7, which is a diagram of the toxic effect of the increased toxic T cells targeting WT1 antigen on human pancreatic cancer cells and tumorigenic cancer stem cells after the dendritic cells that can target cancer stem cells and their combinations with T cells, and the dendritic cells co cultured with WT1 antigen acquire dendritic cells that can target WT1 antigen ability, and then inject dendritic cells that can target WT1 antigen ability into the body of cancer patients, according to one embodiment of the present disclosure.
[0062] It can be seen from the figure that compared with untrained T cells only, the relative cytotoxicity ability of WT1 targeted T cells trained with WT1 antigen to human pancreatic cancer oncogenic stem cells (TCSC) has increased by about 13 times. The combination of dendritic cells that can target the cancer stem cells with T cells cultured using the exosomes of oncogenic cancer stem cells of human pancreatic cancer as antigen presenters, when its concentration is 30 pg (TCSCexo-30) or 50 pg (TCSCexo- 50. Its cytotoxicity to oncogenic cancer stem cells of human pancreatic cancer has increased by about 6 times and 8 times in sequence compared with the cytotoxic T cells that can target WT1 antigen, so it obviously has better cytotoxicity.
[0063] In addition, compared with untrained T cells, the cytotoxicity of dendritic cells to tumor forming cancer stem cells of human pancreatic cancer in the concentrations of 30 pg and 50 pg, the combination of dendritic cells that can target the cancer stem cells with T cells obtained by using the exosomes of tumorigenic cancer stem cells of human pancreatic cancer as antigen presenters has increased by about 76 times and 102 times in sequence, which is much higher than that of untrained T cells.
[0064] In summary, the dendritic cells targeting cancer stem cells and their combination with T cells provided by the present disclosure can indeed solve the problem of existing immunotherapy being unable to effectively kill cancer stem cells, resulting in tumors having drug resistance, recurrence, or metastasis, making it impossible for tumors to receive effective treatment.
[0065] Those skilled in the art to which the present disclosure belongs can understand from the above content that the present disclosure can be embodied in other specific forms without changing the technical concept or essential features of the disclosed content. In this regard, the illustrative forms disclosed in this article are for illustrative purposes only and should not be interpreted as limiting the scope of the disclosed content. On the contrary, this disclosure is intended to cover not only such illustrative forms, but also various changes, modifications, equivalents, and other forms that may be included within the spirit and scope of the present disclosure as defined in the attached patent application scope.
Claims
What is claimed is:
1. A combination of dendritic cells that can target the cancer stem cells with T cells, prepared by the following steps:SI : separating a plurality of monocytes and a plurality of T cells from a peripheral blood collected from a human body;S2: cultivating the monocytes into a plurality of dendritic cells through induction;S3 : extracting a plurality of cancer stem cell surface antigens from a plurality of cancer stem cells;S4: co-culturing each of the cancer stem cell surface antigens of the cancer stem cells with the dendritic cells to obtain a plurality of dendritic cells that can target the cancer stem cells; andS5: co-culturing the dendritic cells that can target cancer stem cells with the T cells to obtain a combination of dendritic cells that can target the cancer stem cells with T cells.
2. The combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, wherein in the step SI, a monocyte line is CD14+monocyte.
3. The combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, wherein in the step SI, a T cell line is CD8+T cells.
4. The combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, wherein in the step S2, a dendritic cell line is CDllc+dendritic cells.
5. The combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, wherein in the step S3, the cancer stem cell surface antigens are extracted from exosomes or vesicles of the cancer stem cells.
6. The combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, wherein in the step S3, the cancer stem cell surface antigens areextracted by repeatedly freezing and thawing the cancer stem cells.
7. A method for preparing a combination of dendritic cells that can target the cancer stem cells with T cells, comprising the following preparation steps:SI : separating a plurality of monocytes and a plurality of T cells from a peripheral blood collected from a human body;S2: cultivating the monocytes into a plurality of dendritic cells through induction;S3 : extracting a plurality of cancer stem cell surface antigens from a plurality of cancer stem cells;S4: co-culturing each of the cancer stem cell surface antigens of the cancer stem cells with the dendritic cells to obtain a plurality of dendritic cells that can target cancer stem cells; andS5: co-culturing the dendritic cells that can target cancer stem cells with the T cells to obtain a combination of dendritic cells that can target the cancer stem cells with T cells.
8. The preparation method as claimed in Claim 7, wherein in the step SI, a monocyte line is CD14+monocyte.
9. The preparation method as claimed in Claim 7, wherein in the step SI, a T cell line is CD8+T cells.
10. The preparation method as claimed in Claim 7, wherein in the step S2, a dendritic cell line is CDllc+dendritic cells.
11. The preparation method as claimed in Claim 7, wherein in the step S3, the cancer stem cell surface antigens are extracted from exosomes or vesicles of the cancer stem cell.
12. The preparation method as claimed in Claim 7, wherein in the step S3, the cancer stem cell surface antigens are extracted by repeatedly freezing and thawing the cancer stem cells.
13. The use of a combination of dendritic cells that can target the cancer stem cells with T cells as claimed in Claim 1, for eliminating the cancer stem cells to prevent tumor drug resistance, recurrence, and / or metastasis.
14. A dendritic cell that can target cancer stem cells, prepared by the following steps:SI : separating a plurality of monocytes and a plurality of T cells from a peripheral blood collected from a human body;S2: cultivating the monocytes into a plurality of dendritic cells through induction;S3 : extracting a plurality of cancer stem cell surface antigens from a plurality of cancer stem cells; andS4: co-culturing each of the cancer stem cell surface antigens with the dendritic cells to obtain the dendritic cells that can target cancer stem cells.
15. The dendritic cells that can target cancer stem cells as claimed in Claim 14, wherein in the step SI, a monocyte line is CD14+monocytes.
16. The dendritic cells that can target cancer stem cells as claimed in Claim 14, wherein in the step S2, a dendritic cell line is CDllc+dendritic cells.
17. The dendritic cells that can target cancer stem cells as claimed in Claim 14, wherein in the step S3, the cancer stem cell surface antigens are extracted from exosomes or vesicles of the cancer stem cells.
18. The dendritic cells that can target cancer stem cells as claimed in Claim 14, wherein in the step S3, the cancer stem cell surface antigens are extracted by repeatedly freezing and thawing the cancer stem cells.
19. A method for preparing dendritic cells that can target the cancer stem cells, comprising the following preparation steps:SI : separating a plurality of monocytes and a plurality of T cells from a peripheral blood collected from a human body;S2: cultivating the monocytes into a plurality of dendritic cells through induction;S3 : extracting a plurality of cancer stem cell surface antigens from a plurality of cancer stem cells; andS4: co-culturing each of the cancer stem cell surface antigens with the dendritic cells to obtain a plurality of dendritic cells that can target cancer stem cells.
20. The preparation method as claimed in Claim 19, wherein in the step SI, a monocyte line is CD14+monocyte.
21. The preparation method as claimed in Claim 19, wherein in the step S2, a dendritic cell line is CDllc+dendritic cells.
22. The preparation method as claimed in Claim 19, wherein in the step S3, the cancer stem cell surface antigens are extracted from exosomes or vesicles of the cancer stem cells.
23. The preparation method as claimed in Claim 19, wherein in step S3, the cancer stem cell surface antigens are extracted by repeatedly freezing and thawing the cancer stem cells.