Human breast cancer cell line with high lung metastasis
A high-metastasis technology of human breast cancer cells, applied in the field of microbial animal cell lines, can solve the problems of short incubation period and low rate of spontaneous lung metastasis, and achieve the effects of wide application, short experimental period, and clear genetic background
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Embodiment 1
[0014] Example 1 Screening and establishment of lung metastatic cells
[0015] The cells involved in the present invention are all cultured in Leibovitz's L-15 cell culture medium containing 10% fetal calf serum (FCS). The culture temperature was 37°C, and the gas environment was 5% CO 2 / 95% air, humidity is saturated humidity. When subcultured, the cells were digested with 0.25% trypsin, and subcultured at a ratio of 1:3.
[0016] 1. The first round of screening of lung metastatic cells
[0017] ① Digest MDA-MB-231 cells in the logarithmic growth phase with 0.25% trypsin to make a single cell suspension;
[0018] ②Under sterile conditions, tumor cells were inoculated into the fat pad (mfp) of the left second papilla of 6-week-old BALB / c female nude mice (BALB / c nude mice) with a 1ml syringe or directly injected into the tail vein (i.v. ), the capacity is adjusted to 1×10 7 Cells / 0.1ml / one (mfp) or 2×10 6 Cells / 0.1ml / piece (i.v.);
[0019] ③After 4 weeks of routine fee...
Embodiment 2
[0031] Example 2 Mice Transplanted Tumor and Lung Metastasis Experiment
[0032] 1. BALB / c nude mice
[0033] ①Digest the MDA-MB-231 and MDA-MB-231 / 08-001H cells in the logarithmic growth phase with 0.25% trypsin, and make single-cell suspensions respectively;
[0034] ②Under sterile conditions, use a 1ml syringe to inoculate tumor cells into the fat pad of the left second papilla of nude mice, with an inoculation volume of 1×10 7 Cells / 0.1ml / piece;
[0035] ③Observe the health status of the nude mice and the tumorigenicity of the tumor cells the next day;
[0036] ④After 4 weeks, 6 nude mice were randomly selected from each group, and their lungs were fixed with Bouin's solution for 24 hours, and then soaked in absolute alcohol until the color of the lung tissue recovered and the metastatic lesions were white nodules, and were examined under a dissecting microscope. The number of metastatic nodules in each lung was observed and recorded; some transplanted tumors were excis...
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