Human hepatocellular carcinoma cell line, and establishing method and application thereof
A method for establishing a method and a technology for cancer cells, applied in the field of cell lines, can solve the problems of large differences in biological characteristics of liver cancer and low biodiversity, and achieve the effect of weight loss and stable characteristics
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Embodiment 1
[0042] Preparation of LIXC-012 cells
[0043] SCID mice: 5 SCID mice, male, weighing 16.0±1.0 g, aged 5 weeks, were raised in SPF environment. SCID mice were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.
[0044] A fresh resection specimen of clinical liver cancer was obtained from Shanghai Zhongshan Hospital (male, 71 years old, primary liver cancer, pathological diagnosis: (right lobe of liver) hepatocellular carcinoma, grade II differentiation, nodular cirrhosis of surrounding liver tissue, pulse Tumor embolus in the tube), immediately immersed in pre-cooled sterile HBSS buffer (containing 500 U / mL penicillin G, 500 μg / mL streptomycin sulfate, and 1.25 μg / mL amphotericin B). In the biosafety cabinet, the specimen was washed with fresh sterile HBSS buffer solution, cut into small pieces of 20-50 mg, and punctured and inoculated subcutaneously in the axillae of SCID mice. 30 days after inoculation, small tumor nodules were found under the skin o...
Embodiment 2
[0049] Biological characteristics and application of LIXC-012 cells
[0050] The invention adopts RPMI1640 culture solution containing fetal bovine serum and insulin to cultivate LIXC-012 cells, so that they can grow and pass stably in vitro for a long time. When the cells are passed to more than 20 generations, the cell properties are gradually stabilized, and the relevant biology, genetics and tissue source identification are carried out until the 50th generation has the same stable properties. Through experimental observation and verification, the LIXC-012 cells grown in vitro are epithelial-like. Under an inverted microscope, the cells adhere to the wall, disperse uniformly, vary in size, and have a cobblestone-like shape. The cells have a typical epithelial-like morphology and lose contact growth inhibition. , showing malignant growth. Genetic studies have confirmed that the cells are aneuploid, with severe chromosome number and structural aberrations, consistent with th...
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