动物模型 z-bo 2020-08-04 593

　　[Modeling mechanism] Nude mice are usually subcutaneously inoculated with tumor mass or cells, and the tumor continues to grow. It can simulate the process of human malignant tumors. The tumor formation rate is high, the tumor formation time is uniform, and the tumor size and weight can be easily measured. Basic and clinical research is easy to conduct. However, the growth, invasion and metastasis behavior of tumor implants is not only related to the biological characteristics of the tumor itself, but also related to the tumor implantation site. Subcutaneous inoculation can keep tumor cells away from the microenvironment of tissues and organs. In the experiment, unlike the original patient's malignant tumor, the specific biological behavior of the transplanted tumor is usually heterotopic transplantation. Transplantation is a kind of tumor transplantation that is increasingly recognized by people, and by injecting human tumors into the host organ as the main part of the tumor, a microenvironment similar to that of human tumors can be obtained. I will. Provide a variety of endogenous growth factors that promote tumor growth.

　　[Modeling method]

　　1. Harvest UMR-106 cells in the logarithmic growth phase, digest with trypsin, wash twice with PBS to remove residual serum, and finally use serum-free medium RPMll640 to adjust the cell number and concentration of the cell suspension method, 4 weeks The female BALB/c-nu/nu nude mice were anesthetized with carbamate, the dosage of carbamate was 600μl/g, the local skin was disinfected regularly before inoculation, and the outer layer of the femur was disinfected under the operating microscope . After exposing and lifting the 0.15 mm x 0.15 mm cortex, slowly inject the cell suspension. After the medullary cavity, the bone flap is reset, sealed with bone wax, and the fascia and skin are sutured. The whole process is carried out in a sterile clean room.

　　2. The tissue blocking method is to enter UMR-106 cells into logarithmic growth phase, adjust the cell concentration, and subcutaneously inoculate 2x1,000,000 cells/female nude mice on the back for 4 weeks. Increase to cm x 2.0 cm x 1.5 cm. Take the non-necrotic tumor tissue, place it in PRMI1640 medium, and cut it into tumor tissue pieces with a diameter of about 0.3 mm with ophthalmic scissors. Take a six-week-old female nude mouse, open the outer side of the femur in the same way as above, send a uniform tumor tissue mass to the medullary cavity, and reset and suture the bone flap. Movies usually show a clear growth trend in the third week.

　　[Characteristics of the model] Two weeks after the tumor tissue mass was implanted or the cell suspension was inoculated, the local bones began to swell, the surrounding tissues were compressed, and the skin turned purple. , Swollen veins. X-ray film showed bone destruction 3-4 weeks after implantation, surrounding soft tissue infiltration, and pathological fractures 6 weeks later. Under an optical microscope, the transplanted tumor cells have different sizes and morphologies, with large nuclei, the ratio of nuclei is reversed, and the nuclei are distributed in nests.

　　[Assessment and application of the model] The advantage of this model is that it can simulate the original growth microenvironment of osteosarcoma, with less tumor avascular necrosis, high lung metastasis rate and dense metastasis. It is mainly used to study the biological characteristics of tumor cell proliferation and invasion, local imaging of osteosarcoma and salvage treatment of limbs; it is used for targeted therapy carrier and chemical sensitivity screening research.