Novel liver cancer biotherapy method
A technology for liver cancer and liver cancer cells, applied in the field of biomedicine, can solve the problems of inability to prevent tumors and little effect
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Embodiment 1
[0023] Example 1 Construction of H22 mouse liver cancer animal model
[0024] H22 ascites tumors were frozen in our laboratory. We inoculated the H22 ascites tumors frozen in liquid nitrogen into the peritoneal cavity of Kunming mice. After 2 weeks, the peritoneal cavity of Kunming mice was covered with H22 ascites. The ascites tumor of H22 was inoculated into the armpit of Kunming mice, and after 2 weeks, it could grow into a solid transplanted tumor of H22 liver cancer.
[0025] Take the H22 ascites tumor fluid, count the cells in the ascites, and dilute to 7.5×10 6 1 / ml and inoculated 0.2ml dilution in the armpit of 10 female 20-22g Kunming mice, and another 10 healthy growing mice were used as controls. Ten days later, the two groups of mice were removed from the eyeballs to collect blood, and dissected to record the tumor tissue, liver, brown adipose tissue (Brown Adipose Tissue, hereinafter referred to as BAT) in the scapula and white adipose tissue (White Adipose Tissu...
Embodiment 2
[0035] Excision model of embodiment 2 brown adipose tissue
[0036] 40 female Kunming mice weighing 20-22 g were anesthetized by injecting pentobarbital sodium solution at 50 mg / kg, and 20 of them had the brown adipose tissue in the scapula removed, and the wound was sutured immediately; the other 20 were sham Surgical treatment, that is, suturing directly after the incision of the scapula. Three days later, 10 mice with brown adipose tissue excised and 10 sham-operated mice were inoculated with H22 tumor fluid in the armpit. Ten days later, the eyeballs of all 40 mice were removed to collect blood, and dissected to record the weights of tumor, liver, brown adipose tissue in the scapula and white adipose tissue in the groin. The results are shown in Table 3. At the end of the experiment, the collected blood was centrifuged, and the supernatant was drawn into a clean EP tube. This was the serum, which was stored in a -80°C refrigerator.
[0037] We found that the weight of tu...
Embodiment 3
[0052] Example 3 H22 mixed primary brown adipocyte (BrownAdipose Cell, hereinafter referred to as BAC) xenograft tumor model
[0053] First, cells from mouse brown adipose tissue were extracted. Take 3-4 week old Kunming mice, kill them by pulling their necks, and soak them in alcohol for 5 minutes. Take out the brown adipose tissue in the scapula, trim it as clean as possible in the ultra-clean workbench, remove the surrounding white adipose tissue, cut it into pieces and place it in the digestive solution (1mg / ml collagenase Ⅰ, 123mM NaCl, 5mM KCl, 1.3mM CaCl 2 , 5mM glucose, 4% BSA, and 100mM Hepes, pH7.4) for half an hour, take the filter to filter, leave the upper undigested tissue to continue digestion, and centrifuge the lower digestion solution to resuspend the cells in the pellet in DMEM-F12 medium containing 20% FBS. After overnight culture, brown fat cells were counted, mixed with H22 cell suspension, and each mouse was injected with 0.2ml of the two mixed suspe...
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