Construction method of novel brain metastatic tumor animal model

A construction method and technology of brain metastases, applied in the field of construction of new animal models of brain metastases, can solve the problems of unfavorable tumor progression, increased modeling mortality, invasion of trachea, etc., to avoid tumor cell leakage and reduce the risk of modeling Mortality, the effect of avoiding thrombosis

Pending Publication Date: 2021-07-23
GUANGDONG GENERAL HOSPITAL
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Problems solved by technology

However, the common carotid artery in mice is relatively thin, with a diameter of no more than 1mm. Even if an insulin needle (0.3mm in diameter) is used, the damage to the blood vessel caused by puncture is very significant, which often leads to massive bleeding after the needle is pulled out. Combining the use of hemostatic materials to compress hemostasis is helpful. It is good for hemostasis, but it also increases the incidence of carotid artery thrombosis, causes cerebral infarction and seizures in mice, and increases the mortality rate of the model. Its ischemic and hypoxic environment is also not conducive to the progression

Method used

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  • Construction method of novel brain metastatic tumor animal model
  • Construction method of novel brain metastatic tumor animal model
  • Construction method of novel brain metastatic tumor animal model

Examples

Experimental program
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Embodiment

[0038] Embodiment, the construction method of a kind of novel animal model of brain metastases

[0039] A method for constructing a novel brain metastases animal model, comprising the following steps:

[0040] 1. Digest and collect tumor cells stably overexpressing luciferase in an ultra-clean workbench, wash once with PBS, count the cells, add PBS buffer to resuspend, and prepare 10 4 / μL concentration of cell suspension, stored in ice bath for later use;

[0041] 2. Intraperitoneally inject 0.2mL / 10g body weight of tribromoethanol solution with a concentration of 1.25%, anesthetize the experimental mouse, fix it on the animal operating table after the anesthesia is satisfactory, fully expose the neck skin, and shave the hair. Clear airway secretions.

[0042] 3. Disinfect the surgical area with 75% alcohol. Make a longitudinal incision in the middle of the neck, up to 5mm below the incisors, down to 5mm above the sternum, separate the two lobes of the thyroid gland, exp...

experiment example

[0046] Experimental example, pathological examination and in vivo fluorescence imaging verification experiment

[0047] A total of 41 models were created using the construction method of Example 1, including 3 strains of mice (male C57BL / 6 mice, female Balb / c mice, female and male Balb / c nude mice) and 5 kinds of cell lines (small mouse breast cancer cell line 4T1, mouse lung cancer cell line LLC, human breast cancer cell line MDA-MB-231, human lung cancer cell line NCI-H2030 and PC-9), it was found that the mouse model constructed by the present invention passed According to in vivo fluorescence imaging observation, the tumor formation rates of the above five cell lines 10 days after injection were: 4T1: 76.47% (13 / 17), LLC: 71.4% (5 / 7), MDA-MB-231: 14.3% (1 / 17). 7), NCI-H2030: 20% (1 / 5), PC-9: 60% (3 / 5). Observed by HE staining of pathological sections, the incidence of metastases in brain tissue sections of the above five cell lines 30 days after injection were as follow...

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Abstract

The invention relates to the technical field of animal model construction, and discloses a construction method of a novel brain metastatic tumor animal model. The construction method disclosed by the invention comprises the following steps: separating a left common carotid artery and a left common carotid artery branch of the neck of a mouse; rotating the head of the mouse, lifting up a common carotid artery traction silk suture, and then temporarily blocking the common carotid artery; lifting up the telecentric end of the external carotid artery, and leading tumor cell suspension into an internal carotid artery towards the proximal end; and after completing leading in the tumor cell suspension, closing an external carotid artery, and opening the common carotid artery. According to the construction method, the pathophysiological process of human tumor brain metastasis is well simulated, the bleeding amount and the probability of tumor cell leakage in the modeling operation are reduced, the modeling success rate is increased, and the death rate of a model is reduced; and the prepared brain metastasis tumor animal model avoids metastasis of multiple organs, the whole-body tumor load of the mouse is reduced, and the mouse model prepared by the construction method can be used for researching the metastasis mechanism and tumor microenvironment of the brain metastasis tumor.

Description

technical field [0001] The invention relates to the technical field of animal model construction, in particular to a method for constructing a novel animal model of brain metastases. Background technique [0002] Currently, the animal model of brain metastases is constructed by injecting the tumor cell suspension into the arterial blood circulation of the mouse through a specific route, so that the tumor cells are delivered to the capillaries in the brain tissue through the blood flow, and finally the tumor cells penetrate the blood. BBB colonization in the brain parenchyma, or by direct intracranial implantation, caused experimental mice to form metastases. The existing injection methods include: 1. left ventricle injection, 2. common carotid artery injection, and 3. stereotaxic intracranial orthotopic injection. The left ventricle injection relies on experience or under the guidance of B-ultrasound, punctures the left ventricle through the intercostal space of the mouse, ...

Claims

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Application Information

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IPC IPC(8): A61D1/00A61D7/00
CPCA61D1/00A61D7/00
Inventor 杨勇周东薛渊吕广钊周爱冬毛瑞王力周立婷
Owner GUANGDONG GENERAL HOSPITAL
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