Tumor cell-induced angiogenesis model experiment
Tumor cell-induced angiogenesis model experiment
The experimental model of tumor cell-induced angiogenesis can be used to (1) establish an in vitro experimental model to facilitate further research, and (2) observe the growth characteristics of tumors as well as their specificities.
Operation method
Cell Culture Technology
Principle
Both primary and secondary tumors are angiogenic once they grow more than 1-2 mm in diameter. This is due to the fact that tumor cells themselves can secrete a variety of growth factors that induce angiogenesis.
Materials and Instruments
Tumor Cells Move 1. Tumor cells in logarithmic growth phase were washed with DEM culture solution, and the cells were collected and re-suspended in DEM culture solution. Caveat The mechanism of angiogenesis is complex, and there are numerous factors that participate in and promote angiogenesis. The number of macrophages in the EMT peritoneal fluid is significantly increased, and their secretion of TNF-α and IL-8 promotes the proliferation of vascular endothelial cells, transforming growth factor-β (TGF-β), platelet-derived endothelial cell growth factor (PD-ECGF), acetylheparinase, angiopoietin (ans), osteogenin (OPN), cyclooxygenase (COX-2), hypoxia-inducible factor-1, laminin (LN), and Osteopoietin (OPN), cyclooxygenase (COX-2), hypoxia-inducible factor-1, laminin (LN), placental growth factor (PLGF), survivin, and erythropoietin (Epo) are all involved in the process of EMT angiogenesis. Common Problems Tumor angiogenesis is an extremely complex process, which generally includes steps including degradation of vascular endothelial stroma, endothelial cell migration, endothelial cell proliferation, endothelial cell pipetting and branching to form vascular rings, and formation of new basement membrane. Tumor angiogenesis occurs partly because tumor cells release angiogenic factors to activate vascular endothelial cells and promote endothelial cell proliferation and migration, and partly because endothelial cells paracrine certain vascular growth factors to stimulate tumor cell growth. The interaction between tumor cells and endothelial cells occurs throughout the process of tumor angiogenesis. Usually, tumor neovascularization occurs as an extension of existing blood vessels in a process similar to typical wound healing and embryogenesis. These new blood vessels provide nutrients to the infiltrating and growing primary tumor, and in turn, the tumor cells secrete a variety of substances to accelerate the formation of tumor neovascularization during the growth process. For more product details, please visit Aladdin Scientific website.
DEM Sodium alginate NaCl EDTA CaCl2 Tepan Blue Formaldehyde
Centrifuge Spectrophotometer Gamma counter
2. Suspended tumor cells were mixed with 1.2% sodium alginate solution, diluted to 0.8% sodium alginate and a cell concentration of (1~5) x106 cells/ml.
3. Tumor cell dilution of 0.8% sodium alginate was loaded into a sprayer, and forcefully pressed the liquid out of the nozzle.
4. When water-soluble sodium alginate is mixed with 80 mmol/l CaCl2, Na+ is replaced by Ca2+ to become a gel.
5. The air pressure and the speed of spraying determine the size of the sodium alginate pellet, which can be rinsed with culture medium or 0.15 mol/ NaCl.
6. Add 0.5 ml of 0.9% NaCl containing 1% EDTA to 0.5 ml of sodium alginate-embedded cells.
7. EDTA breaks the binding bond between Ca2+ and alginate, turning the gel into liquid.
8. Tumor cells were determined by Taipan blue.
9. Embedded cells were diluted with 50% sodium alginate solution in order to obtain the desired number of tumor cells in a constant volume.
10. Tumor cells encapsulated in 200 ul of sodium alginate solution were injected subcutaneously with a 21-gauge needle along the white line of the abdomen, using the sodium alginate solution as a control.
11. After 3~21 days, the animals were executed and the alginate pellets were isolated subcutaneously from the abdomen:
(1) 10% neutral formaldehyde fixation, paraffin embedding, sectioning, staining, histochemistry and immunochemistry.
(2) Zhuge weighed them, soaked and shaken in room temperature downloaded water overnight, and the hemoglobin content in the supernatant was determined by Drabkin's analytical method at 540 nm on a spectrophotometer.
(3) The 51Cr-labeled erythrocytes were injected into the mice from the tail vein 1 h before the animals were executed.
(4) After pulling the neck to execute the mice, the isolated sodium alginate pellet and a piece of tissue away from the pellet were weighed and the radioactivity was determined by gamma counter.