Method for establishing human tumor xenotransplantation model cultured in vitro

Abstract

The invention relates to the field of human xenotransplantation model research, in particular to a method for establishing a human tumor xenotransplantation model cultured in vitro. The establishing method includes the following steps: (1) collecting a tumor tissue sample; (2) placing a tumor tissue in a culture solution; (3) completing the preparation for low-temperature transportation and treatment; (4) cutting the tumor tissue into grain size in a sterile operating table; (5) cleaning the tumor tissue and then placing the tumor tissue in a hole plate 48 for culture; (6) detecting tissue apoptosis by in situ end labeling method; (7) selecting immunocompromised NCG mice to perform anesthesia and skin treatment; (8) implanting the cultured tumor tissue into the renal capsule of the mice; (9) completing modeling after 3 months. According to the invention, tumor tissue blocks are selected for modeling for the xenotransplantation model, which is beneficial to preserving the histopathological and genetic characteristics of primary tumors; and moreover, the obtained tumor tissue are cultured in vitro, which is beneficial to tumor typing and clinical practice; the addition of PD 98059 and artificial matrix glue can greatly reduce tissue apoptosis, which is beneficial to improving the success rate of transplantation.

Description

technical field [0001] The invention relates to the research field of a human-derived xenograft model, in particular to a method for establishing a human-derived tumor xenograft model cultured in vitro. Background technique [0002] Cancer is a major public health problem worldwide, and its morbidity and mortality are increasing year by year. At the same time, the incidence of cancer is the main cause of death today (Siegel R L et al., 2017); A model that highly simulates the occurrence and development of tumors in the human body is a key step in tumor research; currently, in vivo mouse models are used to simulate the development of tumors in humans, and the currently used in vivo models of mice include genetic engineering mice, chemically induced Models and human-derived xenograft models, among which the human-derived xenograft model (English: patient-derived xenografts, abbreviated as PDX) is based on the inoculation of fresh tumor cells or tissues from patients into immun...

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Problems solved by technology

[0003] According to different transplantation sites, PDX models are mainly divided into subcutaneous transplantation, orthotopic transplantation, and renal subcapsular transplantation. Subcutaneous transplantation is generally located in the bilateral groin or axilla of mice, which is easy to observe and regularly measure tumor size, but the site lacks blood. The interaction between donor and tumor-host, and rarely spread and metastases; orthotopic transplantation is the most suitable transplantation method for the tumor microenvironment, but this model is difficult to operate, the tumor formation rate is low, and it is limited to the mammary fat pad , pancreas, brain and other parts are not universal (Hidalgo M et al., 2014); the renal capsule is rich in blood supply and has a high tumor formation rate, so the renal capsule is an ideal site for transplantation (Cunha G R et al. al., 2016)

[0004] Human-derived xenograft models can be divided into single cell suspension transplantation and tissue block transplantation according to different inoculation materials. The cell block transplantation model is easy to operate and has a high tumor formation rate; however, due to the in vitro screening of the cell block transplantation model, the expression of gene profiles It has changed, lost the heterogeneity within the tumor, and lacked the interaction of the tumor microenvironment, and could not objectively reflect the characteristics of primary tumor occurrence, proliferation, and metastasis (Daniel VC et al., 2009; Siolas, D et al ., 2013); the genome mutations in the tissue block transplantation model were reduced, which more truly reflected the biological characteristics of the primary tumor (Bendavid Uet al., 2017; Loukopoulos P, 2004). The success rate of model transplantation ranges from 23% to 75%. At the same time, the time limit for clinical operations and animal experiments and the location of tumor sites are relatively high. Animals need to be transplanted in a very short time after surgery, and tumors with high activity need to be selected. Nest site transplantation can achieve a high success rate of transplantation, which requires the close cooperation of clinicians, which has become a serious obstacle to the popularization and application of this technology (Siolas D et al., 2013)

[0005] At present, the biggest problem facing the application of PDX technology is that the human-derived xenograft model is mainly based on cell transplantation, which loses the heterogeneity in the tumor, and lacks the interaction of the tumor microenvironment, which cannot objectively reflect the occurrence, proliferation, and development of the primary tumor. The characteristics of metastasis, etc.; as for tissue block transplantation, due to different tumor types and complicated operations, the success rate of transplantation and modeling time vary greatly, which need to be further optimized and improved.

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