A simple method for constructing and preparing a human tumor mouse model

Abstract

The application discloses a simple human tumor mouse model and a preparation method thereof. The preparation method of the human tumor mouse model comprises the following steps: a new-born mouse within 24 hours is given a human tumor whole antigen suspension through nose dropping and intraperitoneal injection, and the same kind of human tumor cells are subcutaneously inoculated when the mouse grows to 5-6 weeks, so as to obtain the human tumor-bearing mouse model. The human tumor-bearing mouse model obtained by the preparation method has normal immune function, can simulate the physiological and biochemical environment of a human being with normal immune function, is especially suitable for screening drugs achieving an antitumor effect by regulating the function of an immune system, and does not have the problem that the tumor nodules are too large to be used, so that the human tumor-bearing mouse model will play a better role in the screening of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a simple method for constructing and preparing a human tumor mouse model. Background Technology

[0002] Cancer is the leading cause of death worldwide. The development of anti-tumor drugs remains one of the most important and popular areas of research in the life sciences and pharmaceutical industry. This development is inseparable from the use of human tumor animal models. Currently, the main method for creating human tumor animal models, both domestically and internationally, is to inoculate human tumor cells or tissues into immunodeficient mice. However, such animal models have a significant drawback: they suffer from immune dysfunction and cannot fully simulate the physiological and biochemical environment of immunocompetent humans. This is especially true for drugs that exert their anti-tumor effects by modulating the immune system; such animal models are simply unsuitable for this purpose. For screening new anti-tumor drugs, animal models carrying human tumors but with normal immune function are the ideal solution.

[0003] In addition, some researchers have successfully obtained mouse models bearing human HepG2 tumors by injecting human liver cancer HepG2 cells into the peritoneal cavity of fetal mice at 16.5 days of gestation, followed by subcutaneous inoculation with human liver cancer HepG2 cells after the fetuses are born. Other researchers have prepared human tumor xenograft models by inoculating Syrian hamsters within 24 hours of birth into the peritoneal cavity and subcutaneously, achieving a lifetime tumor bearing rate of 66.67%. However, both of these methods have drawbacks. Injecting tumor cells during the embryonic stage is a complex and difficult procedure, unsuitable for large-scale production of mouse models bearing human tumors. Inoculating newborn animals with tumor cells results in tumor cell growth as the animals grow, rendering the tumors too large for experimental use before the animals reach adulthood. Summary of the Invention

[0004] To address the above issues, our research team developed and modified the modeling method. We prepared a suspension of human tumor cell suspension using ultrasound to disrupt the cell flow and then administered it via intranasal and intraperitoneal injections to newborn mice within 24 hours. This exposed the mice to the human tumor antigen before their immune systems matured, thus inducing immune tolerance. Once the mice reached adulthood, they were subcutaneously inoculated with the same type of human tumor cells, resulting in a human tumor-bearing mouse model. This human tumor-bearing mouse model exhibits normal immune function, similar to that of immunocompetent humans, but the human tumor nodules are not excessively large, making it more suitable for screening anti-tumor drugs. It also avoids the drawbacks of inoculating human tumor cells during the embryonic or neonatal period. This preparation method is simple to operate and allows for the mass production of human tumor-bearing mouse models.

[0005] Therefore, the first objective of this invention is to provide a method for preparing a human tumor mouse model, comprising the following steps: administering a human tumor whole antigen suspension to newborn mice via intranasal drops and intraperitoneal injection, and then subcutaneously inoculating the mice with the same type of human tumor cells when the mice have grown to 5-6 weeks, thereby obtaining a human tumor-bearing mouse model.

[0006] Furthermore, the preparation steps of the human tumor whole antigen suspension are as follows: culturing human tumor cells in vitro, collecting human tumor cells, and preparing 1×10⁻⁶ cells / mL suspension. 7 Human tumor cell suspension was prepared by ultrasonic disruption of human tumor cell suspension to obtain human tumor whole antigen suspension.

[0007] Furthermore, the administration of human tumor antigen suspension to newborn mice via nasal drops and intraperitoneal injection is specifically as follows: 1 drop of human tumor antigen suspension is administered to one nostril of newborn mice within 24 hours of birth, followed by an intraperitoneal injection of 50-100 μL of human tumor antigen suspension per mouse.

[0008] Furthermore, the dosage of the subcutaneous inoculation with allogeneic human tumor cells is: 5 × 10⁶ per mouse. 5 ~5×10 6 0.2 mL / piece

[0009] A second objective of this invention is to provide the application of the above-described preparation method in the field of animal model construction.

[0010] Furthermore, the animal model is a human tumor-bearing mouse model.

[0011] A third objective of this invention is to provide the application of the human tumor-bearing mouse model obtained by the above-described preparation method in screening drugs for the prevention or treatment of tumors.

[0012] A fourth object of the present invention is to provide a method for screening candidate drugs for the prevention or treatment of tumors, comprising the step of administering a test candidate drug to a human tumor-bearing mouse model obtained according to the preparation method described above.

[0013] The present invention has the following beneficial effects:

[0014] 1. The human tumor-bearing mouse model obtained by the preparation method of this invention has normal immune function and can simulate the physiological and biochemical environment of humans with normal immune function. It is particularly suitable for screening drugs that achieve anti-tumor effects by regulating the function of the immune system. Furthermore, the human tumor-bearing mouse model does not have tumor nodules that are too large to be used, and it will play a better role in the screening of anti-tumor drugs.

[0015] 2. The present invention provides a simple method for producing human tumor-bearing mouse models, and can simultaneously produce large quantities of human tumor-bearing mouse models. Detailed Implementation

[0016] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0017] Example 1: Preparation of a mouse model bearing human glioma

[0018] 1. Human glioma cell line (GFP-U87) was cultured and expanded in vitro. After reaching a sufficient quantity, human glioma cells were collected and prepared into 1×10⁻⁶ cells. 7 Human glioma cell suspension was prepared by sonication to obtain adult glioma whole antigen suspension.

[0019] 2. Twenty mice (Balb / c) born within 24 hours of birth were given human glioma whole antigen suspension via nasal drops (1 drop per mouse) in one nostril, followed by intraperitoneal injection of human glioma whole antigen suspension (50 μL per mouse).

[0020] 3. When the mice reached 5 weeks of age, they were subcutaneously inoculated with the same type of human glioma cells on the right side of the neck and back, 1×10⁶ cells per mouse. 6 0.2 mL / piece

[0021] 4. Seven to ten days later, tumor nodules grew subcutaneously at the sites where human glioma cells were inoculated in mice. The tumor nodules grew to 0.8–1 cm. 3 It can provide screening tests for anti-tumor drugs.

[0022] 5. The success rate of the human glioma-bearing mouse model prepared using the technical solution of the present invention is greater than 85%.

[0023] Example 2: Preparation of a mouse model bearing human lung adenocarcinoma

[0024] 1. Human lung adenocarcinoma cell line (95-D) was cultured and expanded in vitro. After reaching a sufficient quantity, human lung adenocarcinoma cells were collected and prepared into 1×10⁻⁶ cells. 7 Human lung adenocarcinoma cell suspension was prepared by sonication to obtain a whole antigen suspension of adult lung adenocarcinoma.

[0025] 2. Twenty mice (Balb / c) within 24 hours of birth were given human lung adenocarcinoma whole antigen suspension via nasal drops (1 drop per mouse) in one nostril, followed by intraperitoneal injection of human lung adenocarcinoma whole antigen suspension (50 μL per mouse).

[0026] 3. When the mice reached 6 weeks of age, they were subcutaneously inoculated with allogeneic human lung adenocarcinoma cells on the right side of the neck and back, with 5 × 10⁶ cells per mouse. 5 0.2 mL / piece

[0027] 4. Seven to ten days later, tumor nodules grew subcutaneously in mice inoculated with human lung adenocarcinoma cells. The tumor nodules grew to 0.8–1 cm. 3 It can provide screening tests for anti-tumor drugs.

[0028] 5. The success rate of the human lung adenocarcinoma mouse model prepared using the technical solution of the present invention is greater than 70%.

[0029] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A method for preparing a human tumor mouse model, characterized by, The mice within 24 hours after birth were given human tumor whole antigen suspension by one drop per mouse through one side of the nostril, and were given human tumor whole antigen suspension by intraperitoneal injection at 50-100 μL per mouse, the concentration of the human tumor whole antigen suspension was 1 x 10 7 When the mice grew to 5-6 weeks, the same human tumor cells were subcutaneously inoculated, the administration amount of the human tumor cells was 5 x 10 5 ~5 x 10 6 per 0.2 mL, and the human tumor-bearing mouse model was prepared.

2. The production method according to claim 1, characterized by, The preparation steps of the human tumor whole antigen suspension are as follows: culturing human tumor cells in vitro, collecting the human tumor cells, and preparing 1×10 7 cells / mL cell suspension; ultrasonically crushing the human tumor cell suspension to obtain the human tumor whole antigen suspension.

3. Use of the preparation method of claim 1 or 2 in the field of animal model construction.

4. Use according to claim 3, characterized in that, The animal model is a human tumor-bearing mouse model.

5. Use of the human tumor-bearing mouse model obtained by the preparation method of claim 1 or 2 in screening drugs for preventing or treating tumors.

6. A method for screening a candidate drug for prevention or treatment of a tumor, characterized by, comprises administering a test candidate drug to the steps in the human tumor-bearing mouse model obtained by the preparation method of claim 1 or 2.