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Mouse-derived oral squamous cell carcinoma cell strain, and preparation method and application thereof

A technology of squamous cells and cancer cells, applied in the field of cell lines

Pending Publication Date: 2021-08-03
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the lack of existing murine oral squamous cell carcinoma cell lines with tumorigenicity in immune-competent mice, the present invention provides a mouse oral squamous cell carcinoma cell line, preparation method and application thereof

Method used

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  • Mouse-derived oral squamous cell carcinoma cell strain, and preparation method and application thereof
  • Mouse-derived oral squamous cell carcinoma cell strain, and preparation method and application thereof
  • Mouse-derived oral squamous cell carcinoma cell strain, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Preparation method of mouse oral squamous cell carcinoma cell line

[0026] Step 1: Add 4-NQO (100 μg / ml) to the drinking water of C57BL / 6J mice for 20 weeks of induction, then replace with normal drinking water, and continue feeding for 4 weeks. A cauliflower-like lesion grows on the back of the mouse tongue. With invasion, the diagnosis was oral squamous cell carcinoma ( figure 1 ). Use high-temperature and high-pressure sterilized tissue scissors and blades to quickly excise the tumor tissue, soak it in high-sugar DMEM culture medium containing 20% ​​fetal bovine serum and 1% penicillin + streptomycin double antibody, and quickly transfer it to a biological safety cabinet for operation . Wash repeatedly with PBS buffer containing 1% double antibody to obtain mouse tumor tissue;

[0027] Step 2: Cut the above mouse tumor tissue into 1*1*1mm small pieces with sterilized tissue scissors, wash with PBS, add 1 ml of trypsin, and incubate in a 37°C cell cul...

Embodiment 2

[0030] Example 2 Detection and identification of mouse oral squamous cell carcinoma cell lines

[0031] (1) Morphological observation

[0032] The cell shape is polygonal, oval or kidney-shaped, and grows in clusters, with large nuclei and abundant cytoplasm.

[0033] (2) Immunofluorescence staining detection

[0034] Step 1: Inoculate the cells on a cell slide, and when the cell density reaches 40% to 50%, rinse twice with PBS buffer for 5 minutes each time, add 4% paraformaldehyde to fix for 30 minutes, and then rinse again with PBS buffer. Rinse twice, 5 minutes each time;

[0035] Step 2: Add 100 microliters of 3% hydrogen peroxide solution to each slice, in order to block endogenous catalase and reduce non-specific staining, incubate at room temperature for 30 minutes, wash with PBS buffer 3 times, each 5 minute;

[0036] Step 3: Add 100 microliters of 5% goat serum to each tablet and incubate at room temperature for 20 minutes;

[0037] Step 4: Add 100 microliters o...

Embodiment 3

[0047] The establishment of the animal model of embodiment 3 mouse oral squamous cell carcinoma cell lines

[0048] Step 1: Anesthetize the C57BL / 6J mice, depilate the upper back, and prepare the above cells into 1*10 7 cells / 1 ml of cell suspension, each mouse was inoculated with 100 microliters of cells;

[0049] Step 2: Observe the state of the mice twice a week, measure the tumor size of the mice (tumor volume=maximum tumor diameter*minimum diameter*minimum diameter / 2), and weigh the body weight of the mice;

[0050] Step 3: When the tumor reaches 1000mm 2 The mice were euthanized before, and the tumor tissues of the mice were collected, fixed in formaldehyde, embedded in paraffin, and sectioned;

[0051] Step 4: Perform HE and immunofluorescence staining on the slices, see the results Figure 7 , Figure 8 .

[0052] To sum up, observed under an optical microscope, the cells were polygonal or elliptical, growing in clusters and adherent to the wall, with large nuclei a...

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Abstract

The invention discloses a mouse-derived oral squamous cell carcinoma cell strain,and a preparation method and application thereof, and the preparation method comprises the following steps: firstly, adding 4-nitroquinoline-N-oxide into mouse drinking water, inducing mouse oral squamous cell carcinoma, and obtaining mouse oral squamous cell carcinoma tumor tissues; carrying out digestion and primary culture on the mouse tumor tissue to obtain mouse tumor primary cells; selecting a monoclonal cell from the primary cells by adopting a monoclonal screening method; and purifying the monoclonal cells, carrying out continuous passage, and screening out a cell strain to obtain the mouse oral squamous cell carcinoma cell strain. The mouse squamous cell carcinoma cells can form tumors on both immunodeficient nude mice and immune-healthy C57BL / 6J mice, so that the problem that the existing human oral squamous cell carcinoma cell line has poor tumor forming effect on immune-healthy mice is solved; the invention has an important value in establishment of a tumor immune mouse model and research of a subsequent interaction mechanism of a tumor and an immune system.

Description

technical field [0001] The invention relates to a cell line, in particular to a mouse-derived oral squamous cell carcinoma cell line, a preparation method and application thereof. Background technique [0002] Tumor cell lines are commonly used tools in tumor research. They play an important role in the study of the occurrence and development of tumors, the molecular mechanism of metastasis, the screening of potential therapeutic targets, and the evaluation of the efficacy of therapeutic drugs. The human-derived oral squamous cell carcinoma cell line can well retain the genetic background and biological characteristics of the patient's tumor, but due to the immune rejection caused by the difference between species, the human-derived oral squamous cell carcinoma The effect of tumorigenesis is unsatisfactory. If human-derived tumor cell lines are used to form tumors in immunodeficient mice, the important role of the interaction between tumors and the immune system in the occu...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02A01K67/027
CPCC12N5/0693C12N5/0632G01N33/5011A01K67/0271C12N2509/10C12N2509/00C12N2503/02G01N2500/10A01K2207/12A01K2227/105A01K2267/03
Inventor 许可傅友田国才
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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