移植性肿瘤动物模型 z-bo 2020-10-14 362

　　(1) Non-solid tumor cell transplantation model

　　Blood samples of clinical leukemia patients cultured and passaged in the abdominal cavity of mice, bone marrow samples or non-solid tumor ascites samples are equal amounts of balanced salt solutions (PBS, etc.). Or, add physiological saline and mix well, centrifuge and separate tumor cells. When culturing non-solid tumor cells in an in vitro suspension culture, the culture solution containing tumor cells is directly collected and centrifuged to obtain tumor cells. The speed and time of centrifugation are 800-1500/min, 5 minutes. Tumor cells collected from blood samples and bone marrow samples of clinical leukemia patients usually contain red blood cells, and the red blood cells are crushed with a hemolytic agent (0.83% NH4Cl, etc.), and then the cells are centrifuged in a balanced salt solution. Flush to dispose of hemoglobin and debris. After collecting tumor cells from the above-mentioned various sources, adjust their concentration to (1×1000000?1×10000000)/ml in the mouse or abdomen or in the abdomen or tail vein of the mouse with serum-free culture medium or balanced salt solution. . Inject SCID mice. 0.2-0.5 ml for mice. The amount of inoculation depends on the degree of malignancy of the cells. Usually a large amount of tumor cells and low-grade tumor cells are injected into clinical specimens, rather than small doses. Seven to ten days after the inoculation, the mouse abdomen enlarged or tumor cells in the blood were detected.

　　In addition, after successful intraperitoneal inoculation of mice, mouse ascites can be extracted to isolate tumor cells, and then subcutaneously inoculated to establish a solid tumor transplantation model (solid tumor cells below), please refer to Creating a transplantation model.

　　(2) Solid tumor cell transplantation model

　　Clinical tumor tissue or tumor tissue that has been successfully transplanted and transferred to animals should be thoroughly cleaned with balanced salt solution to remove blood stains and surrounding non-tumor tissues should be cleanly cut first, and then finely sliced. (The smaller the incision, the better); completely suspend it in a balanced salt solution, transfer it to a centrifuge tube, and place it at room temperature for 5 minutes; discard the top layer of the lavage solution containing cell debris, and remove the small tumor tissue Leave the precipitate; finally soak and digest: Resuspend the tumor tissue sediment with 5-10 times the amount of tumor tissue digestion solution, and then use a pipette to evenly suck it up. At the same time, place it at 37°C for 30 minutes, shaking it every 5 minutes. Use an equal amount of serum-containing culture medium to complete the digestion. Let it stand at room temperature for 5 minutes, collect the suspension containing the cells in the upper layer, and place it in another centrifuge tube. The smaller, incompletely digested underlying tumor tissue can be submerged and digested again. Collect the tumor cell suspension several times, centrifuge at 800-1000/min for 5 minutes, discard the upper layer of liquid to leave the cell pellet, suspend the cells in the complete culture medium and evenly aspirate the liquid. Perform a cell count and adjust the cell concentration. It exceeds 1 x 10000000/ml, and the animal is eventually vaccinated subcutaneously or in situ. The number of inoculated cells is 1000000-10000000 cells, the amount of subcutaneous inoculation is 0.2-0.3 ml, and the amount of in situ inoculation is 20-500μl. The digestive juice used for immersion digestion is different from tumors of different tissues. Trypsin (PBS) is used to prepare tumor tissues derived from common organs and use it for tumor tissues derived from nerve tissue or stromal tissue (collagenase (serum-free medium) preparation). Usually, it takes about 10 days after inoculation to detect tumor cell growth.