34 results about "Immunodeficient Mouse" patented technology

The invention belongs to the technical field of biotechnology and animal models, and discloses a double-person-derived mouse model for simulating a tumor immune microenvironment and a construction method and application thereof. The method comprises the following steps: firstly, pretreating NSG immunodeficient mice by adopting low-dose whole-body irradiation in combination with double-antibody targeted bone marrow depletion, and transplanting CD34 + hematopoietic stem cells from the same human donor to complete human immune system reconstruction; separating tumor primary cells, tumor-related fibroblasts and tumor vascular endothelial cells of the same donor, performing three-dimensional co-culture to obtain homologous human tumor organs, and performing in-situ inoculation to immune reconstruction mice to obtain a target model. The core defects of MHC mismatching, low immune reconstruction efficiency, poor tumor immune microenvironment simulation degree, low clinical consistency and the like of an existing model are overcome, and the method can be used for tumor immune treatment drug screening, microenvironment mechanism research and personalized tumor treatment scheme verification.

The application discloses application of a CCL15 neutralizing antibody in preparation of a medicine for inhibiting growth of liver cancer, and comprises the following steps: S1, constructing a mouse liver cancer PDX model and performing passage; S2, randomly dividing the immunodeficient mouse after passage into a control group and an experimental group, and performing dosing treatment on the control group and the experimental group every three days; S3, PDX model tumor tissue evaluation, stripping tumor tissues after 11 times of dosing of the control group and the experimental group, and respectively observing and recording the volume and weight of the tumor tissues of the control group and the experimental group; the method for inhibiting growth of liver cancer provided by the application, CCL15 acts on the stellate cells to activate the stellate cells and make the expression spectrum of the stellate cells change significantly, the CCL15 neutralizing antibody has an anti-tumor effect in the liver cancer PDX model through injection, and tumor-derived CCL15 can indirectly promote growth of liver cancer cells by activating the hepatic stellate cells.

The invention relates to an immune system humanized mouse biological system for preparing a fully humanized monoclonal antibody and a method for preparing the fully humanized monoclonal antibody. The method comprises the following steps: using a constructed immune system humanized mouse and a VLP chimeric antigen; the HSC immune system humanized mouse is an immunodeficient mouse transplanted with human immune cells, the human immune cells are reconstructed, and antigen-specific B cells and fully humanized antibodies can be generated in the immune system humanized mouse by using a VLP chimeric antigen without firstly activating DC and antigen-specific T cells. In addition, by coupling a VLP antigen and a target antigen, a specific B cell and a fully human antibody of any target can be generated.

Provided in the present invention is an sgRNA targeting the mouse FLT3 gene. Further provided in the present invention is a method for constructing a mouse having a humanized immune system, the method comprising: injecting an AAV vector comprising a cytokine gene into a severely immunodeficient FLT3-gene-knockout mouse; optionally irradiating the FLT3-gene-knockout mouse; and injecting human hematopoietic stem cells into the FLT3-gene-knockout mouse.

This invention relates to the field of leukemia research, specifically disclosing an animal model of CAR-T therapy for leukemia complicated by cytokine release syndrome (CRS), its preparation method, and its application. The method for preparing the animal model of CAR-T therapy for leukemia complicated by CRS includes: obtaining CD19 CAR-T cells: peripheral blood mononuclear cell isolation, T cell magnetic bead sorting, T cell activation, lentiviral infection, and CAR-T cell expansion; constructing an animal model of CRS after high- and low-dose CAR-T infusion: implanting Nalm-6 cell line into living animals to induce leukemia tumor burden, obtaining living animals with leukemia tumor burden, and injecting CD19 CAR-T cell line into the living animals with leukemia tumor burden. This invention provides a novel application for the animal model of CAR-T therapy for leukemia complicated by cytokine release syndrome. By using severely combined immunodeficient mice and inoculating them with high- and low-burden tumor cells, and infusing different doses of CAR-T cells, this invention constructs an in vivo model that can simulate clinical CRS, providing research guidance for clinically exploring more effective interventions.

The invention discloses primers and a method for detecting Corynebacterium bovis and application of the primers and the method in detection of pathogenic bacteria of immunodeficient mice. The primers comprise an upstream primer and a downstream primer aiming at 16S rRNA (ribosomal Ribonucleic Acid) genes of the Corynebacterium bovis. The invention establishes a qPCR molecular detection method for the pathogenic bacteria Corynebacterium bovis of the immunodeficient mice, and the qPCR molecular detection method has the advantages of strong specificity, high sensitivity, high detection efficiency and the like, and can quickly and accurately detect the Corynebacterium bovis in the immunodeficient mice and the environment. The establishment of the detection method is beneficial to timely discovery and control of the bacterium, and has important significance for guaranteeing the environmental safety of personnel and the health of experimental animals. In future experimental animal quality control and related scientific research work, the qPCR detection method is expected to be widely applied, and powerful technical support is provided for further deep research on biological characteristics and pathogenic mechanisms of corynebacterium bovis.

This invention belongs to the field of biomedical technology, specifically relating to a human-derived cell line for peritoneal metastasis of gastric cancer and its applications. The invention involves collecting ascites from gastric cancer patients and culturing it into organoids. These organoids are then transplanted subcutaneously into immunodeficient mice to form xenografts (PDOX). The transplanted organoids are digested and passaged in vitro to obtain a purified cell line. The primary isolated cell line is then inoculated into the peritoneal cavity of immunodeficient mice to induce the formation of metastatic ascites. Cells are extracted from the metastatic ascites and cultured again into organoids to construct human gastric cancer ascites cells, named BZGC. The cell line constructed in this invention has the ability to form 100% metastatic ascites in the mouse peritoneal cavity, fully simulating the entire process of ascites formation in gastric cancer peritoneal metastasis. The human-derived gastric cancer ascites cells described in this invention provide a stable, reliable, and highly clinically and pathologically similar in vitro and in vivo model for studying the mechanism of gastric cancer peritoneal metastasis, drug screening, and personalized treatment.

The invention discloses a method for implanting a cryopreserved human tissue into a mouse body, which comprises the following steps of: resuscitating the cryopreserved human tissue, implanting the cryopreserved human tissue into an immunodeficient mouse body, continuously feeding the mouse, and adding bFGF (basic fibroblast growth factor) and VEGFA (vascular endothelial growth factor A) when the cryopreserved human tissue is implanted. The invention further discloses a construction method of the cryopreserved tissue and immune system double-person-derived mouse model. The survival rate and structural integrity of the cryopreserved human tissue are remarkably improved by optimizing the size of a transplantation block, performing combined treatment on growth factors and other key technologies, so that the cryopreserved human tissue can be more effectively reconstructed in an immunodeficient mouse body, and a higher level of structural maturity is shown in histology. The two-person-derived mouse model constructed by combining cryopreserved tissues and an immune system can provide a better research platform for vaccine and efficacy evaluation.

A NOD.Cg-Prkdcscid NSG) mouse which is genetically modified such that the NSG mouse lacks functional major histocompatibility complex I (MHC I) and lacks functional major histocompatibility complex II (MHC II) is provided according to aspects of the present invention. According to specific aspects the genetically modified NSG mouse is a NOD.Cg- Prkdcscid H2-K1tml Bpe H2-Ab1em Mvw H2-D1tml H2rg Wjl / SzJ NSG- RIP-DTR (IAnull) mouse, or a NOD.Cg-B2m'm" Une Prkdcscid H2dlAbl-Ea H2rg!"1 Wjl / SzJ (NSG- B2M ull (IA mouse. Human immune cells and / or human tumor cells are administered to a genetically modified immunodeficient mouse according to aspects described herein and assays of one or more test substances can be performed using the provided mice. 20 24 20 47 25 09 J ul 2 02 4 A B S T R A C T 2 0 2 4 2 0 4 7 2 5 0 9 J u l 2 0 2 4

The invention relates to a construction method and application of an immune system sheep-derived mouse model, and the construction method comprises the following steps: 1) taking an adaptively cultured severe immunodeficiency mouse, and injecting busulfan to inhibit the hematopoiesis of the mouse; 2) injecting sheep spleen lymphocytes into the body of the pre-treated immunodeficient mouse in an orbital vein manner; 3) after the sheep cells are transplanted in the step 2), detecting the cell proportions of sheep CD45 +, CD4 + and CD8 + in the peripheral blood of the mouse every week so as to detect the sheep CD45 +, CD4 + and CD8 + in the peripheral blood and / or tissues, and determining that the immune system sheep mouse is successfully constructed if the sheep CD45 +, CD4 + and CD8 + can be continuously detected for 13 weeks; (4) injecting and infecting Brucella into the abdominal cavity of the mouse in the step (3) to obtain the Brucella infected sheep-derived mouse model.According to the method, sheep spleen lymphocytes are transplanted into the mouse body to construct the immune system sheep-derived mouse, and a more effective and more convenient animal model is provided for studying diseases such as Brucella taking sheep as a host.

The invention provides a novel human distal bile duct cancer cell line with TP53 missense mutation and application, the novel human distal bile duct cancer cell line CBC3T-3 is established, the cell line is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C202555), the uniqueness and stability of the cell line are proved through STR typing and karyotype analysis, and the TP53 missense mutation novel human distal bile duct cancer cell line has the advantages that the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3 can be used for preparing the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3; and a plurality of driver gene mutations including TP53 missense mutation are carried. The CBC3T-3 has strong proliferation, invasion and migration capabilities, has high tumor formation rate in immunodeficient mice, is resistant to cis-platinum and sensitive to paclitaxel and gemcitabine, and provides an experimental basis for selection of clinical chemotherapy regimens. According to the model, the TP53 missense mutation type distal bile duct cancer in-vitro model is successfully established, and a key experimental platform is provided for deeply researching the drug resistance mechanism of the TP53 missense mutation type distal bile duct cancer and developing an individualized treatment strategy aiming at the subtype of the TP53 missense mutation type distal bile duct cancer.

This invention discloses a method for constructing a PBMC-derived humanized mouse scleroderma model. Human peripheral blood mononuclear cells are transplanted into immunodeficient mice, and a human immune system is reconstructed in the mice. Bleomycin is injected into the local skin of the mice in a round-point manner to induce fibrosis and form a scleroderma model. This application can more comprehensively simulate the complex pathological process of immune abnormalities and tissue fibrosis coexisting in human scleroderma, and provides a more realistic and systematic experimental platform for in-depth exploration of the immune mechanism of this disease.

Inventors have firstly demonstrated that the transcriptomic signature of PTENpc- / - LSCmed cells is enriched in human MSPC signature, associated to metastases and ADT resistance. Using scRNAseq specifically with sorted PTENpc- / - LSCmed cells, they identified three distinct LSCmed clusters and showed that castration favors the emergence of the most stem-like LSCmed subpopulation by cell plasticity. In the latter, they identified the transcription factor FOSL1 / AP-1 and the kinase PIM1 as relevant therapeutic targets, and they showed that combined inhibition using JQ1, a BET / AP-1 inhibitor, and CX-6258, a pan-PIM kinase inhibitor, efficiently prevents PTENpc / - LSCmed cell growth in organoids. These findings were confirmed in the human prostate HPV10 cell line here identified as a robust model of Club / Hillock cells. Combined drug delivery to castrated PTEN-null mice induced a significant prostate weight decrease associated with the reduction of histopathological phenotypes and dramatically altered organoid-formation capacity of LSCmed cells sorted from these tumors. Accordingly, the drug combination significantly delayed tumor growth of MSPC-like human PC-3 cells subcutaneously injected into castrated immunodeficient mice. Altogether, this work shows a new therapeutic potential of combined FOSL1 and PIM1 targeting to prevent, or at least delay, the growth of MSPC-like cells. Accordingly, the invention relates to i) an inhibitor of FOSL1 and ii) an inhibitor of PIM1, as a combined preparation for use in the treatment of prostate cancer in a subject in need thereof.

The invention discloses a construction method and application of a glioma white matter invasion cell model, and belongs to the field of biomedicine. The method comprises the following steps: constructing a model carrying resistant and fluorescent glioma stem cells (GSC-Luc) through lentivirus transfection, injecting the model into the cranium of an immunodeficient mouse through stereotactic injection, separating tumor core region (GSC-C) and white matter invasion region (GSC-WM) cells, and establishing a paired cell model with the same genetic background. The white matter invasion ability of the model is verified through in-vitro white matter extract culture and in-vivo HE staining, and the model is utilized to reveal that the GSC-WM has higher malignant degree (interstitial score, balling ability and proliferation ability enhancement) and drug treatment resistance (such as TMZ curative effect difference). The model fills the blank of an in-vitro model for glioma white matter invasion research, and provides a new tool for mechanism research and drug development.

A neutralizing antibody capable of potent neutralization of SFTSVs and the use thereof. The neutralizing antibody specifically recognizes amino acids at 26th, 39th, 42nd, 46th, 50th and 53rd sites of the sub-domain I and amino acids at 260th, 263rd, 268th, 271st and 301st sites of the sub-domain III of the extracellular domain of SFTSVGn. The neutralization activity of the neutralizing antibody ZS004-1C5 in inhibiting SFTSV(DBV) infection of half of cells is 67 thousands times that of MAb4-5 which is the only one fully-humanized neutralizing antibody known so far. Simply by means of one inoculation, the neutralizing antibody ZS004-1C5 can prevent STAT1- / - immunodeficient mice from a lethal attack caused by viruses with a 10-fold median lethal dose (LD50), and therefore has the potential to be put into clinical treatment, and may improve the survival rate for patients with a severe fever with thrombocytopenia syndrome (SFTS) and reduce the risk for intensive care.

The application belongs to the technical field of tumor treatment, and discloses a method for enhancing the anti-tumor effect of an oncolytic virus M1, and specifically discloses application of albendazole in preparation of a product for enhancing the anti-tumor effect of the oncolytic virus M1. The application first proposes that ABZ can increase the therapeutic effect of OVM1 in treating tumors, and has small toxicity and low price. It is proved through experiments that OVM1 combined with ABZ treatment can significantly promote the activity inhibition of tumors in vitro, and the anti-tumor effect of the combined treatment on immune-competent mice is far greater than that on immune-deficient mice; further detection finds that this process depends on CD8 + T cells play a synergistic anti-tumor effect. At the same time, ABZ and OVM1 combined with CTLA4 antibody can not only further enhance the anti-tumor effect, but also overcome the drug resistance of ICB treatment based on CTLA4. The application provides a new strategy for synergistic oncolytic virus treatment and overcoming ICB drug resistance.

This invention relates to an animal model of mediastinal lymph node metastasis in lung cancer and its construction method. The core of this invention lies in the precise intrapulmonary in situ microinjection technique to directly implant human lung cancer cells into the lung parenchyma of immunodeficient mice. Utilizing the natural lymphatic drainage pathway of the lung tissue, tumor cells are induced to migrate to the mediastinal lymph nodes, thus successfully constructing a complete metastasis chain model from the primary tumor to regional lymph nodes. This model can more realistically simulate the microenvironment and process of lung cancer lymph node metastasis, and is suitable for metastasis mechanism research, drug evaluation, and the development of personalized treatment strategies, possessing high translational medicine value.

The present application provides a preparation method of a non-human mammal model of type 1 diabetes. Specifically, the present application induces islet beta cell destruction in a non-immunodeficient mouse by using T cells specifically targeting GAD, constructs T cell autoimmune type 1 diabetes in line with the actual pathogenesis, and has application prospects in the field of diabetes research.

The application provides a construction method of a humanized double-sensitization animal model. The PBMCs of a peanut and pollen co-allergic patient are transplanted to an immunodeficient mouse, and a double-allergen sensitization humanized mouse model is constructed by combining the design of peanut sensitization and pollen-induced sensitization model. The application further provides a construction method of a systemic allergic animal model, which comprises the step of sensitizing and exciting the double-allergen sensitization humanized mouse model with crude peanut extract. The double-allergen sensitization humanized mouse model has high molding rate, stable model, no toxicity and harmlessness, and has specificity; the systemic allergic reaction model can be effectively constructed by food allergen excitation, the multiple allergic characteristics of the human body are simulated, the clinical relevance is strong, and important choices and bases are provided for basic research clinicalization.

The present invention relates to a personalized humanized animal model transplanted with peripheral blood mononuclear cells (PBMCs) and a production method therefor, and more particularly, to a personalized humanized animal model generated by administering peripheral blood mononuclear cells (PBMCs) of a donor to an immunodeficient mouse, and a production method therefor, wherein the immunodeficient mouse is established as a personalized humanized subject by administering a compound containing busulfan or cyclophosphamide thereto.

This invention relates to the field of tumor animal model technology, and provides a method for constructing an osteosarcoma PDX model and its application. The method for constructing the PDX model includes the following steps: S1 pretreatment of tumor samples from osteosarcoma patients, and S2 subcutaneous inoculation in mice. S1 specifically includes: first washing the tumor samples with physiological saline, then storing and transporting the tumor samples in a low-temperature culture medium at 0-4°C, and then washing the tumor samples with room-temperature PBS solution; and mechanically cutting the tumor samples into 2-3 mm pieces. 3 The tissue blocks. S2 specifically includes: based on the number of tissue blocks formed from the tumor sample, preparing at least two immunodeficient mice for each patient to construct the PDX model; selecting inoculation sites from both sides of the lower limbs and / or both sides of the upper limbs of each mouse. By establishing tumor-bearing models through multiple sites, the limited number of tumor samples and experimental animals can be utilized to the maximum extent, rapidly expanding the number of PDX models and improving modeling efficiency and cost.

A method of identifying anti-tumor activity of a test substance is provided along with a genetically-modified, immunodeficient mouse and methods of use, wherein the genetically-modified, immunodeficient mouse enables in vivo investigation of the interactions between the human immune system and human cancer.

The present application belongs to the field of biomedical technology, and relates to a construction method of an experimental animal model and a constructed experimental model, in particular to a method for constructing a kidney humanized mouse model based on human cells and a constructed kidney humanized mouse model. The construction method comprises the following steps: injecting mesenchymal stem cells, human umbilical vein endothelial cells and human renal tubular epithelial cells separated from urine into immunodeficient mice subcutaneously or subcapsularly to obtain a humanized kidney vascular unit mouse model communicating with the host. The model construction has the following characteristics: the cells used are derived from urine, have the characteristics of sufficient clinical source and convenient sampling, and the kidney source cells derived from urine can represent a unique individual alone, can be used to establish an individualized kidney humanized mouse model based on patients, and have the potential to be applied to individualized precision medical research; the model can form a functional human kidney vascular unit in mice, and has great application prospects in individualized precision medical treatment of kidney disease and human kidney specific virus infection research.

The present application belongs to the technical field of animal model construction, and aims at the problem of the lack of human hepatic stellate cells leading to the liver fibrosis animal model, and provides a construction method and application of a liver fibrosis mouse model based on a human hepatic stellate cell strain.The present application uses an immunodeficient mouse as an animal model, uses luciferase and red fluorescent protein labeled LX-2, and injects MCT into the abdominal cavity to make a liver injury model of the mouse, injects LX-2 into the spleen after liver injury, transplants human LX-2 cells into the NSG mouse, and then injects CCl4 into the abdominal cavity of the mouse.Through the method, the implantation, distribution, proliferation and liver fibrosis of the transplanted LX-2 in the liver are successfully studied.The model is suitable for human in-vivo experiments of biological treatment and drug treatment and other treatment methods targeting hepatic stellate cells or fibroblasts.

This invention provides the application of GCKR and its inhibitors in the preparation of drugs for treating pancreatic cancer, belonging to the field of biomedical technology. The application of GCKR and its inhibitors in the preparation of drugs for treating pancreatic cancer reveals for the first time that GCKR is a key metabolic switch driving liver metastasis in pancreatic cancer, and that high expression of the GCKR gene can promote the growth and liver metastasis of pancreatic cancer in mice. The proliferation, invasion, and migration abilities of pancreatic cancer cells in the GCKR low-expression group were significantly lower than those in the normal control group. Using a pancreatic cancer orthotopic xenograft model constructed in highly immunodeficient mice, the invention demonstrated in vivo that knocking out GCKR can inhibit the growth of pancreatic cancer, indicating that GCKR inhibitors block the progression of pancreatic cancer by downregulating GCKR; conversely, overexpression plasmids that promote GCKR expression can upregulate GCKR expression and promote pancreatic cancer progression. This provides a new application of GCKR and offers new ideas for the treatment of pancreatic cancer.

This invention discloses a humanized Sirpα The construction methods of immunodeficient mice and their application in human immune reconstitution belong to the field of medical experimental model construction technology. Rag2 ‑ / ‑ IL2rg ‑ / ‑ Using animals with T, B, and NK cell immune dysfunction caused by mutations as the background strain, sgRNA and dsDNA were specifically designed, and their endogenous components were processed using CRISPR / Cas9 gene editing technology. Sirpα The entire genome was modified to be humanized, thus creating a humanized genome. Sirpα Immunodeficient mice. This method is simple to operate, highly reproducible, and constructs a fully humanized mouse. Sirpα Immunodeficient mice exhibit a higher immunodeficient phenotype and can achieve a higher rate of human T cell chimerism after transplantation with human peripheral blood mononuclear cells, while effectively reducing the risk of graft-host disease.

The invention belongs to the technical field of human tumor cell lines, and provides a human lung cancer cell line TRI-LC21 and application thereof, and the preservation number of the cell line is CCTCC NO: C202581. The cell line is successfully constructed through primary culture, purification screening and in-vitro stable passage methods, so that the technical problem that a standardized in-vitro model which is clear in source, stable in genetic background, capable of realizing long-term passage and capable of truly simulating the biological characteristics of SMARCA4 defective chest undifferentiated tumors (SMARCA4-UT) is lacked in the prior art is solved. The cell line has typical SMARCA4-UT pathological characteristics, including complete deletion of BRG1 protein expression and the like, can stably pass more than 30 generations in vitro, and can form tumors in immunodeficient mice. The SMARCA4-UT model is suitable for pathogenesis research, drug screening and development of treatment strategies of SMARCA4-UT.

The invention discloses a mouse model construction method for simulating intravenous injection circulating tumor cells, and belongs to the field of cell model construction. The invention discloses a method for constructing a tail vein injection circulating tumor cell mouse model for simulating lung metastasis of breast cancer. The method comprises the following steps: S1, treating a tumor operation specimen before transplantation; s2, constructing a PDX model mouse; s3, obtaining a circulating tumor cell suspension; s4, transplanting the circulating tumor cell suspension into the renal capsule of the immunodeficient mouse; s5, obtaining a tail intravenous injection circulating tumor cell mouse model for simulating the lung metastasis of the breast cancer; and S6, monitoring, verifying and evaluating the model. The problems that in the prior art, sources of circulating tumor cells are mixed, and deviation exists between the circulating tumor cells and actual conditions in clinic are solved, the goodness of fit between the circulating tumor cells and the clinical metastasis process is greatly improved, the number of metastases can be precisely regulated and controlled, and the reliability of data and the effectiveness of statistical analysis are ensured.

The application provides a method for constructing a double humanized hepatitis B mouse model characterized by T cell immune reconstruction. The method is characterized by promoting humanized T cell reconstruction, and comprises the steps of constructing a human liver chimeric immunodeficient mouse model, infecting the human liver chimeric immunodeficient mouse with HBV, and implanting human immune cells. The results show that hAlb, HBV DNA and HBsAg can be continuously detected in the blood of the constructed mouse, HBsAg and HBcAg can be detected in the liver, and human immune cell infiltration mainly in the form of CD3+ T cells can be detected in the blood, liver and other organs, thereby forming a double humanized hepatitis B mouse model of the liver and the immune system. The humanized mouse model construction technology of the application can be used to study the interaction mechanism of HBV and immune cells in the process of hepatitis B. The technical scheme provides a good animal model for the research of hepatitis B progression, immune cell exhaustion mechanism and clinical hepatitis B drug treatment.