动物造模 z-bio 2023-03-08 452

　　Objective To construct a mouse model of humanized immune system and transplant it into human prostate cancer cell lines, so as to be used for the research of prostate cancer immunotherapy drugs.

　　Methods Mononuclear cells (PBMCs) were isolated from fresh human peripheral blood by Ficoll density gradient centrifugation and injected into NPG mice via tail vein to establish a mouse model with human immune system. At the 3rd, 4th, 5th, and 6th week after injection, the peripheral blood of mice was collected by tail docking method, and the dynamic changes of human CD45++CD3+ T cells in the peripheral blood of mice were monitored by flow cytometry. Mice were subcutaneously implanted with 1 × 10' prostate cancer 2R1 cells at 3 weeks post-injection, and tumor growth was monitored twice a week. When the tumor grew to about 1000 mm², the mice were euthanized; flow cytometry, immunohistochemistry, HE staining and other methods were used to analyze the infiltration of human immune cells in the peripheral blood, spleen and tumor tissue of the humanized mice.

　　RESULTS: High levels of CD45+CD3+ T cells were detected in the peripheral blood of model mice 3 to 6 weeks after transplantation of human PBMCs. The mice were sacrificed at the 6th week. The results of HE staining and immunohistochemical staining showed that the spleen and tumor tissues of the model mice had infiltration of human CD4+ and CD8+ T cells.

　　Conclusion The immunohumanized mouse model of prostate cancer has been successfully constructed, and its peripheral blood, spleen and prostate cancer tumor tissue have high levels of human-derived T cell infiltration, which is a good preclinical model for prostate cancer immunotherapy in the next step. Foundation.