动物造模,药效评价,移植性淋巴瘤 z-bio 2024-09-02 389

　　[Modeling mechanism] Transplanted lymphoma models can be established by implanting established lymphoma cell lines or patient lymphoma tissues into immunodeficient animals, revealing the characteristics of the tumor. It is currently the most studied tumor model. At present, there are two main transplantation methods for transplant lymphoma models both domestically and internationally: cell implantation and tissue block transplantation.

　　[Method of Modeling]

　　1. Cell implantation of human lymphocytes (Raji), 1 × 1000000 per mouse, intraperitoneal injection in 5-6 week old female BALB/c-nu/nu mice, usually results in significant ascites around 22 days. If the number of inoculated cells increases, the time for ascites to appear will be shortened. This model can be continuously passaged. Under sterile conditions, nude mouse transplanted tumors can be prepared into cell suspensions or ascites tumor cells. After centrifugation, primary cells can still be inoculated according to the above method to form ascites.

　　2. In situ implantation of tissue blocks: Fresh tumor tissue blocks were obtained from the primary lesion and liver metastasis of human primary colorectal malignant lymphoma during surgery. Anesthesia was administered to the recipient animals, and an abdominal midline incision was made to identify the colon. A 3mm incision was made deep into the mucosal layer on the serosal surface of the intestinal wall, and a 1 cubic millimeter sized tumor tissue block was implanted into the colonic mucosal layer of naked mice through a 10.0 non-destructive suture line. The tumor tissue block was fixed to the intestinal mucosal layer and sutured with the abdomen. Then, in situ organ screening was performed in vivo, and a 1 cubic millimeter tumor fragment from the same tumor source liver metastasis was transplanted into the colon mucosal layer of another nude mouse. After tumor formation, the liver metastasis was transplanted into the colon mucosal layer of the nude mouse. Repeat 4 consecutive screenings in nude mice. Take liver metastases for colon transplantation in nude mice, continue to raise them after surgery, and observe for 60-90 days. When the tumor bearing nude mice were in a dying state, one of them was euthanized by cervical dislocation surgery, and the transplanted tumor was systematically dissected and removed. Part of the tumor tissue was continuously passaged between mice using primary transplantation method, with 5-10 mice passaged each time; The other part of the tumor tissue is frozen in liquid nitrogen for future use, and relevant indicators are tested. Other tumor bearing nude mice were raised to near death or natural death to observe the growth, invasion, and metastasis of their tumors.

　　[Modeling mechanism] Transplanted lymphoma models can be established by implanting established lymphoma cell lines or patient lymphoma tissues into immunodeficient animals, revealing the characteristics of the tumor. It is currently the most studied tumor model. At present, there are two main transplantation methods for transplant lymphoma models both domestically and internationally: cell implantation and tissue block transplantation.

　　[Method of Modeling]

　　1. Cell implantation of human lymphocytes (Raji), 1 × 1000000 per mouse, intraperitoneal injection in 5-6 week old female BALB/c-nu/nu mice, usually results in significant ascites around 22 days. If the number of inoculated cells increases, the time for ascites to appear will be shortened. This model can be continuously passaged. Under sterile conditions, nude mouse transplanted tumors can be prepared into cell suspensions or ascites tumor cells. After centrifugation, primary cells can still be inoculated according to the above method to form ascites.

　　2. In situ implantation of tissue blocks: Fresh tumor tissue blocks were obtained from the primary lesion and liver metastasis of human primary colorectal malignant lymphoma during surgery. Anesthesia was administered to the recipient animals, and an abdominal midline incision was made to identify the colon. A 3mm incision was made deep into the mucosal layer on the serosal surface of the intestinal wall, and a 1 cubic millimeter sized tumor tissue block was implanted into the colonic mucosal layer of naked mice through a 10.0 non-destructive suture line. The tumor tissue block was fixed to the intestinal mucosal layer and sutured with the abdomen. Then, in situ organ screening was performed in vivo, and a 1 cubic millimeter tumor fragment from the same tumor source liver metastasis was transplanted into the colon mucosal layer of another nude mouse. After tumor formation, the liver metastasis was transplanted into the colon mucosal layer of the nude mouse. Repeat 4 consecutive screenings in nude mice. Take liver metastases for colon transplantation in nude mice, continue to raise them after surgery, and observe for 60-90 days. When the tumor bearing nude mice were in a dying state, one of them was euthanized by cervical dislocation surgery, and the transplanted tumor was systematically dissected and removed. Part of the tumor tissue was continuously passaged between mice using primary transplantation method, with 5-10 mice passaged each time; The other part of the tumor tissue is frozen in liquid nitrogen for future use, and relevant indicators are tested. Other tumor bearing nude mice were raised to near death or natural death to observe the growth, invasion, and metastasis of their tumors.

　　[Model Features]

　　1. The cell implantation model shows the formation of ascites, usually around 10ml. Under the microscope, a large number of tumor cells can be seen, and there are tumor masses in the abdominal cavity, mainly distributed in the mesentery, close to the intestinal wall and lymph nodes. The thickening and adhesion of the mesentery are extremely obvious. The tumor cells are of uniform size, with deep nuclear staining, multiple nuclear divisions, less dense interstitial arrangement, obvious necrosis of tumor nodules, no obvious infiltration in the spleen, follicular proliferation, obvious germinal center, and more megakaryocytes. No tumor cell infiltration was observed in the liver, kidney, or lung. Chromosome examination showed that among all metaphase polyploids, the centromere banding pattern was consistent with that of human chromosomes, which is a characteristic of human chromosomes. The commonly used methods for cell inoculation are intraperitoneal or subcutaneous injection. Generally speaking, models inoculated subcutaneously can have tumor nodules grow faster due to skin laxity, resulting in longer host lifespan and easier research and observation. By injecting formed cell lines, an 8-cell lymphoma peritoneal model capable of continuous passage can be established. The tumor formation rate is 100%.

　　2. The tissue block in situ implantation method, a highly metastatic model similar to human colon malignant lymphoma, not only matches human colorectal malignant lymphoma cells in terms of histopathology, immunohistochemistry phenotype, ultrastructure, DNA ploidy level, and chromosome karyotype, but also has a tumor transplantation growth rate and liquid nitrogen cryopreservation survival rate of 100%.

　　Model Evaluation and Application

　　The transplantable tumor model is the most commonly used animal model for screening anti-tumor drugs. Its advantages lie in easy operation and application of interfering factors, stable tumor formation in animals, high tumor formation rate, small individual differences, little difference in tumor formation time, and the ability to continuously transplant animals of the same species or strain for long-term preservation for experimental purposes. However, this type of tumor has a fast growth rate, high proliferation rate, and short volume doubling time, which is significantly different from human tumors.

　　2. The spontaneous metastasis model is a difficult problem in the research field of human cancer metastasis models. The metastasis model prepared by the in situ implantation method of tissue blocks mentioned above has stable biological characteristics and provides basic experimental materials for studying the spontaneous metastasis of gastrointestinal malignant lymphoma; This method plays an irreplaceable role in the in-depth study of the biological characteristics, tissue development, etiology and pathology, invasion and metastasis mechanisms, experimental treatment, and other aspects of malignant lymphoma of the colon.