Esophageal cancer cell line and application thereof
A technology for cancer cells and esophageal cancer, applied in the fields of biology and oncology, to achieve fast growth, clear background, and good in vitro culture expansion
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Embodiment 1
[0029] A tumor tissue sample was taken from a patient with esophageal cancer (the patient was a 57-year-old female, the tumor was located in the middle of the esophagus, the postoperative pathology showed moderately differentiated squamous cell carcinoma, the TNM stage was T2N0M0, and the patient's clinical stage was 1B.). After the separated fresh esophageal cancer tissue was washed with PBS in a sterile ultra-clean bench, it was cut into pieces and separated into 0.5-1mm pieces with ophthalmic forceps, scissors and other instruments in a petri dish. 3 Spread the small piece on the bottom of the dish, and add 10mL of 10% FBS (GIBCO company), 1% double antibody DMEM / F12 cell culture medium (GIBCO company) to the culture dish, put it in 37 ℃, 5% CO 2 cultured in an incubator. Change the medium after 5 to 7 days, discard the necrotic and floating small pieces in the tissue pieces, and carry out subculture. During the subculture process, the fibroblasts were continuously removed...
Embodiment 2
[0031] The subcultured human esophageal squamous cell line ZEC-118 cells were used to observe the growth of living cells under an optical microscope (Japan OlympusIMT-2 inverted microscope). like figure 1 As shown in the photo of the optical morphology of the cells, it can be seen that the cells are growing vigorously, the background is clear, and impurities are rare. The cells are flat and irregular polygons.
Embodiment 3
[0033] Cell Index refers to the change of electrical impedance generated by the interaction between living cells and the microelectrode in the well of the detection plate. The xCELLigence Cell Function Analyzer converts these signals into specific parameters to become the Cell Index. The cell index is a good measure of the state of the cell - growth, proliferation, shape change, death, stress, etc. The cell index has been reviewed and adopted by many journals.
[0034]Human esophageal squamous cell line ZEC-118 cells in good growth state were taken, digested with trypsin, made into cell suspension, and counted. Add culture medium to the E-Plate detection plate of the xCELLigence cell function analyzer and measure the background impedance value, then add 100 μL cell suspension (5000 cells) to the E-Plate detection plate, and place it in an ultra-clean bench at room temperature for 30 minutes. Put the E-Plate detection plate with added cells on the detection table (the detection...
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