56 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 invention discloses a brain glioma animal model construction method based on an AAV mediated immune microenvironment. The method comprises the following steps: selecting an immunodeficient mouse at the right age; human CD34 + hematopoietic stem cells are injected for immune system reconstruction; human CD45 + immune cells in the peripheral blood PBMC are detected; the method comprises the following steps: selecting tumor tissues with high-expression brain glioma marker genes, carrying out rapid qPCR (quantitative polymerase chain reaction) or targeted RNA-seq immunohistochemical multiple verification, and screening consistent tumor tissues; the method comprises the following steps: physically dicing tumor tissues, treating the tumor tissues with pretreatment liquid, digesting the tumor tissues into single-cell suspension, and inoculating the single-cell suspension into a mouse cranium; an AAV vector carrying a specific immune regulation gene is injected into the tail vein of the mouse, and the brain glioma immune microenvironment is regulated; when it is observed that the tumor tissue grows stably and has immune cell infiltration characteristics, construction of the brain glioma animal model is completed. Compared with an existing model, the brain glioma model which is more stable, more scientific and closer to the human immune background is provided, and the brain glioma model is suitable for related immunotherapy drug screening and mechanism research.

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.

The invention provides a prostate cancer stem cell culture medium, a culture method and application, and relates to the technical field of biomedicine. The additives comprise the following components in the basal culture medium in concentration: 1-3% (v / v) of fetal calf serum, 1 * N2, 1 * B-27, 10-50 ng / ml of EGF, 10-50 ng / ml of bFGF, 1-10 nM of androgen DHT, 20-100 [mu] M of ascorbic acid, 0.02-3 [mu] M of a GSK3 inhibitor, 0.1-20 [mu] M of a TGF-beta inhibitor and 0.1-20 [mu] M of a ROCK inhibitor. The primary prostatic cancer stem cells separated from the prostatic cancer tissue block can be well proliferated in vitro, and the biological characteristics of the prostatic cancer stem cells are reserved. The amplified prostate cancer stem cells can be transplanted into the body of an immunodeficient mouse to construct a tumor xenotransplantation model. Compared with a method for directly transplanting tumor tissue blocks, the method for transplanting the amplified prostate cancer stem cells can improve the xenotransplantation success rate.

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 present invention relates to the fields of scientific research and medicine. Based on a method of xenotransplantation of immune mice, flow cytometry is performed on the bone marrow of recipient mice by means of immunological detection, and surface markers of human and mouse megakaryocytes are simultaneously labeled to eliminate interference from mouse megakaryocyte cells. This allows for more accurate determination of the level of human megakaryocyte hematopoietic reconstruction, establishes an accurate and feasible method for in vitro evaluation of the megakaryocyte reconstruction function of human hematopoietic stem cells, and increases the success rate of future clinical treatments.

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.

The invention discloses a breast cancer organoid transplantation tumor model construction method, and particularly relates to the technical field of breast cancer organoids, and the method comprises the following steps: 1, selecting NOD-SCID mouse immunodeficient mice, avoiding immunological rejection, feeding the mice under the condition of no specific pathogen, and ensuring the healthy state; 2, culturing the human breast cancer cell line in an incubator with the temperature of 37 DEG C, humidification and 5% CO2; 3, collecting the cultured cancer cells by using cold PBS, and quickly suspending the cells in the ice-cold Matrigel; 4, anesthetizing the mouse by using isoflurane to ensure the comfort and safety in the operation process; and 5, implanting the cell suspension into the subcutaneous part of the mouse by using an injector, so that visible tumors can be quickly formed. The breast cancer organ transplantation tumor model can be rapidly constructed, application is convenient, and the actual using effect is good.

The present invention discloses a method for constructing an animal model of glioma based on AAV-mediated immune microenvironment, comprising: selecting immunodeficient mice of appropriate age; injecting human CD34+ hematopoietic stem cells for immune system reconstruction; detecting human CD45+ immune cells in peripheral blood PBMC; selecting tumor tissues that highly express glioma marker genes, performing rapid qPCR or targeted RNA-seq immunohistochemistry multiple verification, and screening tumor tissues that meet the requirements; physically cutting the tumor tissue and treating it with a pretreatment solution, digesting it into a single cell suspension, and inoculating it into the mouse's skull; injecting an AAV vector carrying a specific immune regulatory gene into the mouse's tail vein to regulate the glioma immune microenvironment; when the tumor tissue is observed to grow stably and have immune cell infiltration characteristics, the construction of the glioma animal model is completed. Compared with existing models, the present invention provides a more stable, more scientific, and more human immune background glioma model, which is suitable for related immunotherapy drug screening and mechanism research.

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.

The invention belongs to the field of medicines, and particularly relates to a construction method of an EBV positive immune reconstruction xenotransplantation tumor model. The method comprises the following steps: taking an SPF-level female severe immunodeficiency mouse, adapting to the environment for 7 + / -3 days, then performing caudal vein injection of human peripheral blood mononuclear cells, and subcutaneously transplanting a human EBV positive gastric cancer cell line AGS / EBV to the right armpit of the mouse next day. The EBV positive immune reconstruction xenotransplantation tumor model provided by the invention has remarkable advantages in the aspects of simulating a human immune system, shortening tumor formation time, revealing that EBV has dual functions of'immune reconstruction-tumor promotion ', supporting treatment strategy development and the like, and an efficient and accurate tool and method are provided for research and treatment of EBV related gastric cancer.

The invention belongs to the technical field of tumor animal models, and particularly relates to a construction method and application of a brain glioma PDTX model. The invention formulates a tumor screening method suitable for constructing the brain glioma PDTX model, tumor samples with relatively high GFAP, SOX2, SLC1A3 and OLIG2 gene transcription levels are screened as ideal tumor sources by performing transcriptome sequencing on tumor tissues, and the model is constructed by the novel brain glioma PDTX model construction method provided by the invention. According to the method, the screened brain glioma tissue is transplanted into the immunodeficient mouse, so that the original microenvironment characteristics of the tumor can be effectively reserved, and the success rate of constructing the brain glioma PDTX model is obviously improved.

The application belongs to the technical field of biological medicine, and particularly relates to an NTD polypeptide of NFE2L1 protein and application thereof in preparation of an antitumor drug. The amino acid sequence of the NTD polypeptide monomer is shown as SEQ ID NO:1. The application constructs the polypeptide of the NTD amino acid sequence of NFE2L1, and it is unexpectedly found that the NTD polypeptide has good anticancer function, and it is verified through a tumor-bearing experiment of an immunodeficient mouse that the NTD polypeptide provided by the application has a significant effect of inhibiting tumor growth.

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 invention discloses a mouse-derived gastric cancer organoid as well as an establishment method and application thereof. The mouse-derived gastric cancer organoid is mainly obtained by culturing a mouse gastric adenocarcinoma cell line MKC1 (Mus musculus) in a culture medium. The mouse-derived gastric cancer organ can be used for constructing various types of gastric cancer animal models and screening gastric cancer treatment drugs. The established mouse source gastric cancer organoid is derived from a transgenic mouse, the repeatability of the construction process is high, the passage stability is good, the tumor characteristics of mouse gastric cancer can be well simulated, tumor formation in an immunodeficient mouse body can be achieved, especially tumor formation in a C57BL / 6 mouse body with a sound immune system can be achieved, the metastatic tumor formation capacity is achieved, and the mouse source gastric cancer organoid has a good application prospect. The method is suitable for gastric cancer immune microenvironment and tumor generation and development mechanism research.

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 provides a method for constructing a humanized kidney organoid xenotransplantation model, which relates to the technical field of xenotransplantation model construction, and comprises the following steps: acquiring physical sign data of a number of immunodeficient mice to obtain a physical sign data set; matching each physical sign data in the physical sign data set with reference physical sign data respectively; after successful matching, using the corresponding immunodeficient mouse as an experimental carrier; pre-processing the experimental carrier to acquire a first image in real time; determining whether the renal capsule of the experimental carrier is exposed by matching the first image with a first reference image; after successful matching, generating a transplantation instruction; after receiving the transplantation instruction, a pipette sucks the kidney organoid and implants the kidney organoid into the renal capsule of the experimental carrier, and after implanting the kidney organoid, suturing the incision; acquiring a second image in real time; determining whether the incision of the experimental carrier is sutured by matching the second image with a second reference image; after successful matching, generating a monitoring instruction.