CN113528445B - PDX modeling adjuvant and application thereof - Google Patents
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Abstract
The invention relates to a PDX modeling adjuvant and application thereof, which comprises transforming growth factor beta, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, insulin, hydrocortisone, Y-27632, fetal bovine serum and solvent. The PDX modeling adjuvant disclosed by the invention is used for improving the inoculation success rate of a PDX model, accelerating the growth of tumors to adapt to experimental requirements, combining various angiogenesis promoting factors and nutrients, solving the problem of difficult establishment of early vascular networks of the PDX model, and combining local anti-inflammatory medicines hydrocortisone and a compound ROCK inhibitor Y-27632 for promoting the growth of stem cells, so that the survival of the tumor stem cells is guaranteed, the apoptosis of the tumor cells is reduced, the modeling success rate of the PDX model is improved, and the growth speed of a tumor allograft model is accelerated.
Description
Technical Field
The invention relates to the field of anti-tumor drug research and development, in particular to a PDX modeling adjuvant and application thereof.
Background
Cancer has long been a major affliction that jeopardizes human health, and many early stage cancer patients have no obvious symptoms and cannot be found until late stage or metastasis, and the treatment results are often disappointing. Although basic research, clinical research and transformed medicine of tumors have achieved a number of achievements, the new treatment methods do not bring ideal treatment regimens and prognostic effects to patients. In order to study tumor development more deeply and to find better methods of drug treatment, it would be of great value to study models of tumors, particularly animal models.
Traditional xenograft tumor models are xenograft (cell derived xenograft, CDX) of human tumor cell lines, i.e. human tumor cells are screened in vitro, subjected to in vitro subculture, established into stable cell lines, and then inoculated into models established by subcutaneous, kidney capsule or in situ transplantation of immunodeficient mice, and the establishment of the models and the application of the models in tumor research have long history. Early in the 90 s of the 20 th century, the national cancer institute (National Cancer Institute, NCI) introduced a "treatment-oriented" drug screening strategy based on 60 cancer patient tumor cell lines derived from 9 different types of tumors (brain, colon, leukemia, lung, melanoma, ovarian, renal, breast and prostate), simply by first performing high-throughput in vitro drug screening with human tumor cell lines and then performing in vivo validation with CDX model. However, researchers gradually find that the biological behavior, gene profile expression level and tumor heterogeneity of human tumor cell lines have larger differences from those of original tumor tissues after long-term in vitro culture, so that the clinical efficacy is not ideal. Studies have shown that only about 1/3 of the drugs identified and screened by this model are validated in the second-phase clinical trial. The reason is that the tumor cell lines of continuous passage adapt to the environment of an external culture dish, lack of tumor microenvironment, and the tumors formed by the cell lines after being planted into an immunodeficiency mouse have homogeneity with the mice, lose the characteristics of primary tumors and cannot objectively respond to the condition of the primary tumors.
The human tumor allograft model (PDX model) is a substitute model widely applied in the field of anti-tumor drug research and development at present. The model is to transplant the tumor tissue of the cancer patient to the immunodeficiency animal, and make the tumor tissue continuously grow and passaged, thereby producing a large number of tumor-bearing animals carrying the tumor of the patient. Since the PDX model maintains the pathology, molecular characteristics and heterogeneity of the patient's tumor, its sensitivity to anti-tumor drugs is the same as that of the donor patient's tumor. Compared with the traditional cell line transplantation model (cell-derived xenograft, CDX), the PDX model is not subjected to in vitro culture, so that the hereditary property and heterogeneity of the primary tumor are well maintained, and the experimental result has better clinical predictability, so that the model is the most excellent tumor animal model at the present stage. The PDX model can be subjected to passage amplification, and researches show that the passage tumor tissue can keep high consistency with the initial tumor tissue, and good conditions are provided for drug screening and research on the disease mechanism of patients. Thus, detecting the efficacy of alternative treatment regimens prepared for clinical application on the PDX model enables prediction of the efficacy of these treatment regimens in the clinical treatment of tumor patients, thereby providing a reliable reference for the physician to select an appropriate treatment regimen. At present, the consistency of the PDX model for clinical administration to patients has been widely accepted.
However, the current PDX model modeling method is still a method for inoculating tumor blocks into mice subcutaneously, the success rate of the method is limited, a large number of unsuccessful modeling situations are often caused, the problems of slow growth, necrosis while growth, unstable growth, incapability of rapid growth expansion and the like are frequently caused, and not only is a large cost expenditure generated, but also the requirement of patient individuation medical model modeling cannot be met. Therefore, the PDX model has low modeling success rate and long period, and is a main cause of unsuccessful current accurate medical experiments.
Disclosure of Invention
Based on this, it is necessary to provide a PDX modeling adjuvant that can improve modeling success rate and modeling speed.
A PDX modeling adjuvant comprising transforming growth factor beta, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, insulin, hydrocortisone, Y-27632, fetal bovine serum, and a solvent.
In one embodiment, the PDX modeling adjuvant has a concentration of the transforming growth factor beta of 100 ng/mL-1. Mu.g/mL, the epidermal growth factor of 10 ng/mL-0.1. Mu.g/mL, the basic fibroblast growth factor of 10 ng/mL-0.1. Mu.g/mL, the vascular endothelial growth factor of 10 ng/mL-0.1. Mu.g/mL, the insulin of 50. Mu.g/mL-0.5 mg/mL, the hydrocortisone of 50. Mu.g/mL-0.5 mg/mL, the Y-27632 of 10. Mu.M-50. Mu.M, and the fetal bovine serum of 5% -15% by volume.
In one embodiment, in the PDX modeling adjuvant, the concentration of the transforming growth factor beta is 100 ng/mL-200 ng/mL, the concentration of the epidermal cell growth factor is 10 ng/mL-20 ng/mL, the concentration of the basic fibroblast growth factor is 10 ng/mL-20 ng/mL, the concentration of the vascular endothelial growth factor is 10 ng/mL-20 ng/mL, the concentration of the insulin is 50 μg/mL-100 μg/mL, the concentration of the hydrocortisone is 50 μg/mL-100 μg/mL, the concentration of the Y-27632 is 10 μM-20 μM, and the volume percentage of the fetal bovine serum is 5% -10%.
In one embodiment, the PDX modeling adjuvant has a concentration of 100ng/mL of transforming growth factor beta, 10ng/mL of epidermal growth factor, 10ng/mL of basic fibroblast growth factor, 10ng/mL of vascular endothelial growth factor, 50 μg/mL of insulin, 50 μg/mL of hydrocortisone, 10 μM of Y-27632, and 5% by volume of fetal bovine serum.
In one embodiment, the PDX modeling adjuvant further comprises penicillin-streptomycin diabody with the concentration of 1X-5X.
In one embodiment, the concentration of the penicillin-streptomycin diabody in the PDX modeling adjuvant is 1X to 2X.
In one embodiment, the solvent is one or more of phosphate buffer, 1640 medium, and DMEM medium.
The invention also provides application of the PDX modeling adjuvant in preparing a PDX animal model.
In one embodiment, the animal used in preparing the PDX animal model is an immunodeficient mouse.
The invention also provides a preparation method of the PDX animal model, which comprises the following steps: PDX modeling adjuvants as described above were mixed with tumor cells and then inoculated into immunodeficient animals.
Through researches, one of the main problems that the modeling success rate of the PDX model is low is that after human tumor cells are inoculated under the skin of an immunodeficiency animal, the human tumor cells are insufficient in nutrition in a short time and cannot adapt to the internal environment of the immunodeficiency animal. Thus, improving the environment of the inoculation site of immunodeficient animals is a major approach to improving the success rate of PDX model inoculation. After tumor tissue or cells are inoculated into an immunodeficient animal, the pro-angiogenic factors secreted by the tumor cells attract the host's vascular network to provide blood supply to the host for adequate nutrition. However, since a certain time is required for establishing a vascular network, the growth of tumor cells causes a higher demand for nutrient and blood oxygen supply, and until the establishment of the vascular network is not reached, cells in the tumor are subjected to apoptosis, calcification or necrosis due to insufficient supply, so that the primary PDX model grows slowly or dies. In addition, the local inflammatory reaction caused by the inoculation of exogenous substances can recruit a large amount of mouse macrophages, and the infiltration of the large amount of macrophages can interfere with the growth of tumor cells, so that the success rate of modeling of a PDX model is reduced, and therefore, the application of the local anti-inflammatory medicine can be helpful for improving the success rate of modeling of the tumor.
The PDX modeling adjuvant disclosed by the invention is used for improving the inoculation success rate of a PDX model, accelerating the growth of tumors to adapt to experimental requirements, combining various angiogenesis promoting factors and nutrients, solving the problem of difficult establishment of early vascular networks of the PDX model, and combining local anti-inflammatory medicines hydrocortisone and a compound ROCK inhibitor Y-27632 for promoting the growth of stem cells, thereby being beneficial to ensuring the survival of the tumor stem cells and reducing the apoptosis of the tumor cells, so that the situation that a tumor graft is trapped in the growth and apoptosis simultaneously is avoided, the growth retardation of the tumors is broken, the modeling success rate of the PDX model is improved, and the growth speed of a tumor allograft model is accelerated.
Drawings
FIG. 1 is the average tumor formation time of mice model in example 1 using different adjuvants PDX;
FIG. 2 is a graph showing the change in body weight of mice model with different adjuvants PDX in example 2;
FIG. 3 is a graph showing tumor growth curves of mice model for PDX using different adjuvants in example 2;
FIG. 4 is a graph showing the change in body weight of mice model for PDX using different adjuvants in example 3;
FIG. 5 is a graph showing tumor growth curves of mice model for PDX using different adjuvants in example 3;
FIG. 6 is a graph showing the change in body weight of mice model for PDX using different adjuvants in example 4;
FIG. 7 is a graph showing tumor growth curves of mice model for PDX using different adjuvants in example 4;
FIG. 8 is a graph showing the change in body weight of mice model PSX using different adjuvants in example 5;
FIG. 9 is a graph showing tumor growth in mice model for PDX using different adjuvants in example 5;
FIG. 10 is a graph showing the change in body weight of mice model PSX using different adjuvants in example 6;
FIG. 11 is a graph showing tumor growth curves of mice model for PDX using different adjuvants in example 6.
Detailed Description
The present invention will be described more fully hereinafter in order to facilitate an understanding of the present invention, and preferred embodiments of the present invention are set forth. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The PDX modeling adjuvant comprises transforming growth factor beta (TGFb), epidermal Growth Factor (EGF), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VFGF), insulin, hydrocortisone, Y-27632, fetal bovine serum and a solvent.
Through researches, one of the main problems that the modeling success rate of the PDX model is low is that after human tumor cells are inoculated under the skin of an immunodeficiency animal, the human tumor cells are insufficient in nutrition in a short time and cannot adapt to the internal environment of the immunodeficiency animal. Thus, improving the environment of the inoculation site of immunodeficient animals is a major approach to improving the success rate of PDX model inoculation. After tumor tissue or cells are inoculated into an immunodeficient animal, the pro-angiogenic factors secreted by the tumor cells attract the host's vascular network to provide blood supply to the host for adequate nutrition. However, since a certain time is required for establishing a vascular network, the growth of tumor cells causes a higher demand for nutrient and blood oxygen supply, and until the establishment of the vascular network is not reached, cells in the tumor are subjected to apoptosis, calcification or necrosis due to insufficient supply, so that the primary PDX model grows slowly or dies. In addition, the local inflammatory reaction caused by the inoculation of exogenous substances can recruit a large amount of mouse macrophages, and the infiltration of the large amount of macrophages can interfere with the growth of tumor cells, so that the success rate of modeling of a PDX model is reduced, and therefore, the application of the local anti-inflammatory medicine can be helpful for improving the success rate of modeling of the tumor.
The PDX modeling adjuvant disclosed by the invention is used for improving the inoculation success rate of a PDX model, accelerating the growth of tumors to adapt to experimental requirements, combining various angiogenesis promoting factors and nutrients, solving the problem of difficult establishment of early vascular networks of the PDX model, and combining local anti-inflammatory medicines hydrocortisone and a compound ROCK inhibitor Y-27632 for promoting the growth of stem cells, thereby being beneficial to ensuring the survival of the tumor stem cells and reducing the apoptosis of the tumor cells, so that the situation that a tumor graft is trapped in the growth and apoptosis simultaneously is avoided, the growth retardation of the tumors is broken, the modeling success rate of the PDX model is improved, and the growth speed of a tumor allograft model is accelerated.
In a specific example, in the PDX modeling adjuvant, the concentration of transforming growth factor beta is 100 ng/mL-1 mug/mL, the concentration of epidermal cell growth factor is 10 ng/mL-0.1 mug/mL, the concentration of basic fibroblast growth factor is 10 ng/mL-0.1 mug/mL, the concentration of vascular endothelial growth factor is 10 ng/mL-0.1 mug/mL, the concentration of insulin is 50 mug/mL-0.5 mg/mL, the concentration of hydrocortisone is 50 mug/mL-0.5 mg/mL, the concentration of Y-27632 is 10 mug-50 mug, and the volume percentage of fetal bovine serum is 5% -15%.
In a specific example, in the PDX modeling adjuvant, the concentration of transforming growth factor beta is 100 ng/mL-200 ng/mL, the concentration of epidermal cell growth factor is 10 ng/mL-20 ng/mL, the concentration of basic fibroblast growth factor is 10 ng/mL-20 ng/mL, the concentration of vascular endothelial growth factor is 10 ng/mL-20 ng/mL, the concentration of insulin is 50 mug/mL-100 mug/mL, the concentration of hydrocortisone is 50 mug/mL-100 mug/mL, the concentration of Y-27632 is 10 mug-20 mug, and the volume percentage of fetal bovine serum is 5% -10%.
In one specific example, in the PDX modeling adjuvant, the concentration of transforming growth factor beta is 100ng/mL, the concentration of epidermal growth factor is 10ng/mL, the concentration of basic fibroblast growth factor is 10ng/mL, the concentration of vascular endothelial growth factor is 10ng/mL, the concentration of insulin is 50 μg/mL, the concentration of hydrocortisone is 50 μg/mL, the concentration of Y-27632 is 10 μM, and the volume percentage of fetal bovine serum is 5%.
In a specific example, penicillin-streptomycin diabodies at concentrations of 1X to 5X are also included in the PDX modeling adjuvant. Preferably, the concentration of penicillin-streptomycin diabody in the PDX modeling adjuvant is 1X-2X.
In a specific example, the solvent is one or more of phosphate buffer, 1640 medium, and DMEM medium.
The invention also provides application of the PDX modeling adjuvant in preparation of a PDX animal model. In one specific example, the animal used in preparing the PDX animal model is an immunodeficient mouse, but is not limited thereto.
The preparation method of the PDX animal model in an embodiment of the invention comprises the following steps: PDX modeling adjuvant as described above was mixed with tumor cell suspension and then inoculated into immunodeficient animals.
In conclusion, the PDX modeling adjuvant of the invention aims to improve the success rate of inoculation of a PDX model, accelerate tumor growth to adapt to experimental requirements, combine various angiogenesis promoting factors and nutrient substances, solve the problem of difficult establishment of early vascular networks of the PDX model, and combine local anti-inflammatory drugs hydrocortisone and a compound ROCK inhibitor Y-27632 for promoting stem cell growth, thereby being beneficial to ensuring the survival of tumor stem cells and reducing the apoptosis of tumor cells, avoiding the situation that a tumor graft is trapped in growth and apoptosis simultaneously, breaking the growth retardation of tumors, further improving the modeling success rate of the PDX model and accelerating the growth speed of a tumor allograft model.
The following are specific examples.
Example 1
After tumor cells were lysed into single cells, counted and resuspended in DMEM. Tumor cells were resuspended in DMEM medium, and then Matrigel (v: v=1:1) was added to the cell suspension, and immediately after the addition of the adjuvant (cell suspension: adjuvant (v: v) =10:1), the cell suspension was transported on ice to the experimental facility for inoculation, and the total amount of inoculated cells was 5×10e5 to 1×10e7 cells/cell. The control group and the experimental group were inoculated with different adjuvants, and the formulation was as follows:
as shown in fig. 1, the following analysis of the modeling situation of the PDX model established from 11 months in 2020 to 5 months in 2021 shows that the average tumor formation time is 92±9 days in 12 PDX models inoculated without the adjuvant, and the average tumor formation time is 27±4 days in 24 PDX models inoculated with the adjuvant, which shows that the adjuvant can significantly accelerate the speed of establishing the PDX model for the tumor of the patient.
Example 2
The established PDX model tumor is collected from tumor-bearing mice and transferred to the next generation animals through cell digestion treatment. I.e. after the tumor had been destroyed into single cells, the cells were counted and resuspended in DMEM. After tumor cells were resuspended in DMEM medium, matrigel (v: v=1:1) was added to the cell suspension, and immediately after mixing with adjuvant, the cells were transported on ice to the experimental facility for inoculation, the total amount of inoculated cells was 5×10e5 to 1×10e7 cells/cell. The cells were aliquoted into 2 groups at inoculation, each with different adjuvants, the formulation was as follows:
as a result, as shown in FIG. 2, no significant change in the body weight of the mice was observed, and the tumor volume reached 500mm 3 The time of (3) days in advance as shown in figure 3 shows that the adjuvant has the effect of accelerating the growth of the PDX model and has no adverse effect on animal health.
Example 3
The established PDX model is thawed by liquid nitrogen, and after cells are thawed, different adjuvants are respectively added and inoculated according to the method, and the formula is as follows:
the results are shown in fig. 4, and the body weight of the experimental group was not significantly different from that of the control group. As shown in FIG. 5, the average tumor volume of the experimental group reached 1510.3.+ -. 487.53mm by 25 days after inoculation 3 Is significantly larger than the 846+/-139.1 mm of the control group 3 The addition of the adjuvant can obviously accelerate the recovery process of the PDX model, and has no adverse effect on animal health.
Example 4
And (3) recovering the established PDX model after freezing the liquid nitrogen, respectively adding different adjuvants after recovering cells, inoculating 9 groups of animals according to the method, testing the influence of different components on tumor growth, and analyzing the tumor promotion difference of the mixed adjuvants and the single component. The adjuvant formulation is shown below:
after 31 days of inoculation, the experiment was ended and no significant change in body weight was found for each group as shown in figure 6. Tumor volumes were measured twice weekly starting at week 2, the tumor growth curves are shown in fig. 7, and the individual tumor composition conditions are shown in the following table.
The results show that each component has a certain effect of promoting the growth of the tumor, and the Cheng Zuo agent prepared by the combination can play a role of obviously more strongly promoting the growth of the tumor.
Example 5
In the study of adjuvant formulations, we have also found that not all cytokines promote, and some of the cytokines even react to, PDX model neoplasia. Such as AZD-5069, developed by AstraZeneca, inc., is orally administered, is capable of selectively antagonizing chemokine receptor (CXCR 2), has reversibility and time temperature dependence, and has an IC50 of 0.79nmol/L. The agent antagonizes CXCR2, thereby preventing tumor cells from attracting chemotactic macrophages. In normal patients, the blocking effect reduces infiltration of tumor chemotactic macrophages, and simultaneously, causes the increase of inhibitory T cells and the inhibition of angiogenesis, thereby having a certain effect of inhibiting tumor growth. However, in the case of immunodeficient animals, the lack of T cells is not expected to result in the theoretical inhibition of tumor by AZD5069, which in turn reduces the ability of chemotaxis of the site of inoculation to attract macrophages and reduces the local inflammatory response. However, this effect is not reflected in practical experiments. For another example, IL-10 (interleukin 10), IL-10 is a well-defined immunosuppressive molecule that inhibits the innate and acquired immune functions of APCs, resulting in immunosuppression. IL-10 is also a growth factor for certain tumor cells, and numerous studies have demonstrated that many malignant tissues or cells can produce IL-10, such as malignant melanoma, colorectal cancer, ovarian cancer, lung cancer, glioma, and the like. Interleukin 10 affects the growth and differentiation of many hematopoietic cells in vitro and can inhibit the function of macrophages as well as T cells. In IL-10 deficient mice, most animals are slow and anemic, although lymphocyte development and antibody response are normal, and suffer from chronic enterocolitis.
After tumor cells were lysed into single cells, counted and resuspended in DMEM. After tumor cells were resuspended in DMEM medium, matrigel (v: v=1:1) was added to the cell suspension, and immediately after mixing with adjuvant, the cells were transported on ice to the experimental facility for inoculation, the total amount of inoculated cells was 5×10e5 to 1×10e7 cells/cell. The control group and the experimental group were inoculated with different adjuvants, and the formulation was as follows:
as a result, as shown in fig. 8 and 9, in the case of adding IL-10 or AZD-5069 when constructing the PDX model, the effect of promoting tumor growth was not shown at day 31, and the tumor volume of the test group was rather smaller than that of the control group. Therefore, not all growth factors favorable for tumor cells can promote the PDX model to form tumors so as to improve the modeling success rate and the modeling speed.
In conclusion, the PDX modeling adjuvant provided by the invention can obviously accelerate the growth of a PDX model in the modeling, passaging and resuscitation processes, so that experimental failure caused by unstable growth of the model is reduced, experimental cost is reduced, and the usability of the PDX model is improved. The adjuvant formula of the invention not only can supplement cytokines required by tumor cells in PDX modeling, remove the growth retardation of the tumor cells, but also can add related components for inhibiting local immune reaction, reducing host immune attack, and maintaining the survival of tumor stem cells and reducing apoptosis, thereby better supporting the growth of the PDX model, solving the problems of long growth cycle and unstable tumor growth of the PDX model and having positive significance for improving the modeling of the related model.
Example 6
After tumor cells were lysed into single cells, counted and resuspended in DMEM. After tumor cells were resuspended in DMEM medium, matrigel (v: v=1:1) was added to the cell suspension, and immediately after mixing with adjuvant, the cells were transported on ice to the experimental facility for inoculation, the total amount of inoculated cells was 5×10e5 to 1×10e7 cells/cell. Different adjuvants are adopted during inoculation, and the formula is as follows:
as a result, as shown in fig. 10 and 11, the higher dose of adjuvant addition did not show a significant tumor growth promoting effect compared to the optimum formulation when constructing the PDX model, whereas the adjuvant was added below the concentration range proposed by the present invention, and the tumor growth was significantly lower than that of the adjuvant group added at the optimum formulation. Higher doses of adjuvant will bring unnecessary experimental costs, while lower doses will not fully exert the adjuvant-added effect, thus it can be seen that experimental group 1 is the optimal dose for the presently selected adjuvant dose.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (8)
1. A PDX modeling adjuvant comprising transforming growth factor β, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, insulin, hydrocortisone, Y-27632, fetal bovine serum, and a solvent; the concentration of the transforming growth factor beta is 100 ng/mL-200 ng/mL, the concentration of the epidermal cell growth factor is 10 ng/mL-20 ng/mL, the concentration of the basic fibroblast growth factor is 10 ng/mL-20 ng/mL, the concentration of the vascular endothelial growth factor is 10 ng/mL-20 ng/mL, the concentration of the insulin is 50 mug/mL-100 mug/mL, the concentration of hydrocortisone is 50 mug/mL-100 mug/mL, the concentration of the Y-27632 is 10 mug-20 mug, and the volume percentage of the fetal bovine serum is 5% -10%.
2. The PDX modeling adjuvant of claim 1, wherein in the PDX modeling adjuvant, the concentration of the transforming growth factor β is 100ng/mL, the concentration of the epidermal growth factor is 10ng/mL, the concentration of the basic fibroblast growth factor is 10ng/mL, the concentration of the vascular endothelial growth factor is 10ng/mL, the concentration of the insulin is 50 μg/mL, the concentration of hydrocortisone is 50 μg/mL, the concentration of the Y-27632 is 10 μΜ, and the volume percentage of the fetal bovine serum is 5%.
3. The PDX modeling adjuvant of claim 1, further comprising penicillin-streptomycin diabody at a concentration of 1X-5X.
4. A PDX modeling adjuvant according to claim 3, wherein the concentration of the penicillin-streptomycin diabody in the PDX modeling adjuvant is 1X-2X.
5. The PDX modeling adjuvant of any one of claims 1 to 4, wherein the solvent is one or more of phosphate buffer, 1640 medium, and DMEM medium.
6. Use of a PDX modeling adjuvant according to any one of claims 1 to 5 in the preparation of a PDX animal model.
7. The use according to claim 6, wherein the animal used in the preparation of the PDX animal model is an immunodeficient mouse.
8. The preparation method of the PDX animal model is characterized by comprising the following steps of: mixing the PDX modeling adjuvant of any one of claims 1-5 with tumor cells and then vaccinating the immunodeficient animal.
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