Construction method of improved human nasopharyngeal cancer in-situ transplantation tumor model

A technology of human nasopharyngeal carcinoma and construction method, which is applied in the field of medical models, can solve the problems of short survival time, low success rate, low survival time acquisition transfer rate, etc.

Inactive Publication Date: 2019-12-06
JINAN UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

All three models have certain disadvantages. The carcinogen induction method has a long cycle and a low success rate, so it is rarely used
This method describes the in situ inoculation method through the oral cavity through the soft palate mucosa to the nasopharynx of nude mice, but during our research, we found some shortcomings in making this model: under the condition of liquid anesthesia, nude mice immediately suffocated after inoculation Death cases are very common; the survival time of nude mice after implantation is short, about 6-14 days, and the short survival time results in a very low acquisition transfer rate

Method used

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  • Construction method of improved human nasopharyngeal cancer in-situ transplantation tumor model
  • Construction method of improved human nasopharyngeal cancer in-situ transplantation tumor model
  • Construction method of improved human nasopharyngeal cancer in-situ transplantation tumor model

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Experimental program
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Effect test

Embodiment 1

[0026] 1 Materials and methods

[0027] 1.1 Cell line and cell digestion solution (0.25% trypsin-EDTA, Gibco), DMEM (containing 10% fetal bovine serum and 1% double antibody solution), PBS buffer, cell incubator, inverted microscope, cell counting plate.

[0028] 1.2 20 BALB / C male nude mice, SPF grade, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., the mice are 5-6 weeks old and weigh 16-18g. Randomly divided into 10 in the control group and 10 in the model group.

[0029] 2 Construction of animal models

[0030] Nasopharyngeal position of nude mice Figure 1A And implant anatomical positions such as Figure 1B Shown.

[0031] 2.1 Preparation of tumor source

[0032] After the human nasopharyngeal carcinoma cell line 5-8F-luc transfected with bioluminescent protein was cultured and passaged, the tumor cells in the logarithmic growth phase were used to prepare a single cell suspension, and the cell concentration was adjusted to 1.4×10 5 / ml, keep cell via...

Embodiment 2

[0048] 1 Materials and methods

[0049] 1.1 Cell line and cell digestion solution (0.25% trypsin-EDTA, Gibco), DMEM (containing 10% fetal bovine serum and 1% double antibody solution), PBS buffer, cell incubator, inverted microscope, cell counting plate.

[0050] 1.2 20 BALB / C male nude mice, SPF grade, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., the mice are 5-6 weeks old and weigh 16-18g. Randomly divided into 10 in the control group and 10 in the model group.

[0051] 2. Construction of animal models

[0052] 2.1 Preparation of tumor source

[0053] After the human nasopharyngeal carcinoma cell line 5-8F-luc transfected with the bioluminescent protein was cultured and passaged, the tumor cells in the logarithmic growth phase were used to prepare a single-cell suspension.

[0054] 2.2 How to make the model

[0055] The specific construction method of the model group is the same as in Example 1.

[0056] In the control group, the injection needle was us...

Embodiment 3

[0060] 1. Materials and methods

[0061] 1.1 Cell line and cell digestion solution (0.25% trypsin-EDTA, Gibco), DMEM (containing 10% fetal bovine serum and 1% double antibody solution), PBS buffer, cell incubator, inverted microscope, cell counting plate.

[0062] 1.2 20 BALB / C male nude mice, SPF grade, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., the mice are 5-6 weeks old and weigh 16-18g. Randomly divided into 20 rats in the control group and 20 rats in the model group.

[0063] 2. Construction of animal models

[0064] 2.1 Preparation of tumor source

[0065] After the human nasopharyngeal carcinoma cell line 5-8F-luc transfected with the bioluminescent protein is cultured and passaged, the tumor cells in the logarithmic growth phase are used to prepare a single cell suspension. The specific method is the same as that described in the content of the invention.

[0066] 2.2 How to make the model

[0067] The specific construction method of the model ...

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Abstract

The invention relates to a nasopharyngeal cancer transplantation tumor model used for the technical field of medical models. The nasopharyngeal cancer transplantation tumor model is improved on the basis of a naked mouse in-situ transplantation tumor model, and aims at overcoming the technical difficulties that in the transplantation process, the incidence of death from suffocation is high, the survival time after transplantation is short, and long-time experiments are not suitable. Inhalation anesthesia is adopted in tumor model preparation, firstly, naked mice reach the anesthesia depth through gas anesthesia, then 105 stable transfection Luc 5-8F cell suspension is sucked through a 1-ml insulin needle, passes through mouth cavities to pass through the position below border white lines of hard palates and soft palates by 5 mm, and penetrates through mucosae to left-side pharyngeal recesses or right-side pharyngeal recesses to be inoculated, the inoculum size is controlled within 25 microliters, and results show that the incidence of nasopharyngeal cancer in the model mice is 100%, and the naked mice dying from dyspnea after transplantation do not occur in the transplantation process. The model is the in-situ nasopharyngeal cancer model which can be used for researching the development and metastasis of in-situ nasopharyngeal cancer, and meanwhile is also suitable for long-term experimental treatment and diagnostic research of nasopharyngeal cancer.

Description

Technical field [0001] The present invention relates to the field of medical models, in particular to an improved method for constructing an orthotopic transplantation tumor model of human nasopharyngeal carcinoma. Background technique [0002] At present, the domestic and foreign animal models of nasopharyngeal carcinoma include: carcinogen induction method, tumor cell transplantation method and transgenic animal model. All three models have certain shortcomings. Among them, the carcinogen induction method has a long cycle and a very low success rate, so it is rarely used. The cost of genetic modification method is too high, the success rate is low, and it is not suitable for long-term large-scale experiments. [0003] Tumor cell transplantation is currently the most commonly used method, but the currently widely used animal models of nasopharyngeal carcinoma are mainly subcutaneous transplanted tumors, but subcutaneous transplanted tumors are easy to form envelopes, are not easy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61D1/00
CPCA61D1/00
Inventor 张水兴黄文慧
Owner JINAN UNIVERSITY
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