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
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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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