108 results about "Immunodeficient Mouse" patented technology

A hierarchy of endothelial colony forming cells (EPCs) was identified from mammalian cord blood, umbilical vein and aorta. A newly isolated cell named high proliferative potential—endothelial colony forming cell (HPP-ECFC) was isolated and characterized. Single cell assays were developed that test the proliferative and clonogenic potential of endothelial cells derived from cord blood, or from HUVECs and HAECs. EPCs were found to reside in vessel walls. Use of a feeder layer of cells derived from high proliferative potential-endothelial colony forming cells (HPP-ECPCS) from human umbilical cord blood, stimulates growth and survival of repopulating hematopoietic stem and progenitor cells. Stimulation of growth and survival was determined by increased numbers of progenitor cells in in vitro cultures and increased levels of human cell engraftment in the NOD/SCID immunodeficient mouse transplant system.

The invention relates to isolated hemangioblast cells. Hematopoietic and endothelial cells are postulated to be derived from a common progenitor, hemangioblast. While hemangioblast has been isolated retrospectively during embryonic stem cell differentiation, it has not been isolated from embryos or from bone marrow. Prospectively stable clonal cell lines have been isolated from mammalian embryos, from embryonic stem cells and from mammalian bone marrow that can differentiate in vitro into tubular structures with both endothelial and hematopoietic markers such as CD34, CD31, Flk-1, TIE2, P-selectin, Sca-1, thy-1, CD45, and smooth muscle actin. Gene expression profiles in the undifferentiated and differentiated cells were consistent with endothelial and hematopoietic differentiation potential. Transplantation studies in isogenic or immunodeficient mice demonstrated that these cells were not tumorigenic. In an appropriate microenvironment, the cells incorporate into the vasculature and participate in hematopoiesis.

The invention relates to a construction method and application of an immunodeficient mouse animal model, belonging to the field of animal gene engineering and genetic modification. Il2rg gene knockout mice can be obtained by using a CRISPR/Cas9 technology; the Il2rg gene knockout mice have an immunodeficient phenotype. The construction method can be widely applied to multiple fields such as humanized animal models as well as stem cell and tumor research.

The invention provides a human tumor stem cell line T3A-A3, which is obtained by separation of liver cancer tissue excised in an operation of a primary hepatic carcinoma patient. The separated cells can be in long-term passage culture in vitro, can grow rapidly and retain stem cell property after more than 100 times of passage. The cell can express markers of multiple stem cells, has self-updatingcapability of stem cells and directional differentiation potentials for different tumor cells, and also has tumor property, tumorigenicity ability and metastatic ability. The invention is a powerfultool for preparing stem cell drugs for targeting tumor, due to the facts that the cells can be in long-term passage culture in vitro and retain unchanged stem cell property, and the tumorigenicity ability and metastatic ability thereof are strong in immunodeficient mice.

The invention relates to a circulating tumor cell mouse model and a construction method and application thereof. The method includes the following steps of 1, subcutaneous transplantation, wherein a primary tumor tissue sample is transplanted into the body of an immunodeficient mouse to construct a primary cancer heterotransplantation model PDX; 2, collection of circulating tumor cells, wherein the circulating tumor cells in peripheral blood in the primary cancer heterotransplantation model obtained in the step 1 are collected; 3, renal sac membrane transplantation, wherein the circulating tumor cells collected in the step 2 are transplanted into the renal sac membrane of the immunodeficient mouse, and then the circulating tumor cell mouse model is successfully constructed. According to the method, the mode of transplantation with two or more times of passages is adopted, the CTCs in the primary cancer heterotransplantation model are transplanted into the body of the immunodeficient mouse through renal sac membrane, and the circulating tumor cell mouse model is obtained; the circulating tumor cell mouse model can be applied to research on the in-vivo metastasis mechanism and proliferation condition of CTCs.

Provided are a B cell-derived iPS cell generated using a convenient technique, a technology for providing a human antibody at low cost using the iPS cell, an immunologically humanized mouse prepared using cells differentiated from the iPS cell, and the like. Also provided are a cloned cell obtained by contacting a B cell with nuclear reprogramming factors excluding C/EBPα and Pax5 expression inhibiting substances, particularly nucleic acids that encode Oct3/4, Sox2, Klf4 and c-Myc, wherein the cloned cell has an immunoglobulin gene rearranged therein and possesses pluripotency and replication competence (B-iPS cell). Still also provided are a method of producing a monoclonal antibody against a specified antigen, comprising recovering an antibody from a culture of B cells obtained by differentiating a B-iPS cell derived from a B cell immunized with the specified antigen, and a method of generating an immunologically humanized mouse, comprising transplanting to an immunodeficient mouse a human immunohematological system cell obtained by differentiating a B-iPS cell.

The invention provides an ACE2 cell humanized mouse model as well as a construction method and application thereof. The mouse model adopts an immunodeficient mouse as a parent and the body of the mouse contains humanized cells for overexpressing an ACE2 receptor. The mouse model can overexpress the ACE2 receptor in human cells in the body of the immunodeficient mouse, the infection efficiency of acoronavirus taking the ACE2 receptor as an infection medium is improved, meanwhile, xenograft rejection can be avoided, and background pollution is reduced. In addition, the mouse model can also be used for transplantation and immune reconstruction of human immune cells, and is of great significance for further research on the killing effect of the human immune system on the virus, screening of vaccines and evaluation of immune treatment means.

The present invention relates to a cervical intraepithelial neoplasia model and the method of establishing model, which is characterized in that the cervical intraepithelial neoplasia tissue is embedded in the immunodeficiency mice to establish cervical intraepithelial neoplasia model. The method of establishing model is characterized in that the cervical intraepithelial neoplasia tissue is respectively taken from the biopsy under the vaginoscope and the tissues which are confirmed by the department of pathology and is inoculated subcutaneously in the back of the mice; the immunodeficiency mice of the present invention can overcome the xenoislet immune rejection reaction and receive the transplantation of the human tumor tissues, the tissue model after the transplantation has significant and stable character, short observation period and greater clinical reference significance of the experimental results. The present invention can explore the mechanism of the carcinogenesis of cervical carcinoma by outcome of the pathological process of CIN I, CIN II and CIN III animal models. Furthermore, by observing the period of the outcome of the cervical intraepithelial neoplasia tissue of the animal model, the present invention provides the feasible clinical animal experimental model for reversing the malignant transformation of the CIN I and CIN II lesion tissues.

Relating to the technical field of aesthetic and plastic surgery, the invention discloses a method for reconstitution of hair follicle in vivo. The method includes the steps of: firstly performing cell separation, culturing and amplifying freshly isolated cells, and using cell spheres or cell lumps for culture, conducting cell preparation before transplantation, directly taking freshly isolated skin single cells, or cultured and amplified epidermal and dermal cells mixed in proportion, or cell spheres or cell lumps, and resuspending them in a culture solution, narcotizing an immunodeficient mouse, using a syringe needle or cutisector to pierce a hole on the skin of the naked mouse, using a pipette gun head to transfer the well prepared cell suspension into the pierced hole, after transferring all the cells, performing covering with a silica gel membrane and conducting suturing, then laying a layer of vaseline gauze, conducting bandaging and timely observation and treatment, thus finding hair growth 3 weeks later. The method provided by the invention has the characteristics of simple operation, small trauma, high hair follicle formation efficiency, and large injectable cell number range, is suitable for injection of clump cells and a large number of cells, and is suitable for development and application in clinical practice or cosmetic medicine for treatment of hair loss diseases.

The invention discloses a method for constructing an ovarian cancer transplantation tumor model based on an organoid method, and belongs to the field of medicines. According to the invention, an "organoid" culture technology is adopted, and a micro-carrier is used as a support material for culturing a patient-derived ovarian cancer cell-3D material complex; then the cell-3D material complex is directly inoculated under the skin of a normal immune mouse, and a patient-derived ovarian cancer transplantation tumor model is constructed, so that a problem that immune-deficiency mice such as severecombined immunodeficiency (SCID) mice, nude mice and the like are adopted in existing human ovarian cancer transplantation tumor models, the price is high, breeding is difficult, the tumor forming rate is low and the like can be solved, especially a problem that the existing immune-deficiency mouse human ovarian cancer transplantation tumor model cannot reflect impotent effects of an organism immune system in occurrence and development processes of tumors is solved, and a problem that an immune-deficiency mouse human transplantation tumor model is long in tumor forming period and cannot meet drug allergy requirements of patients urgently needing clinic drug therapy is solved. The method is applied to the field of mouse model construction.

The invention provides a preparation method and application of a transgenic mouse with severe immunodeficiency. The method comprises the step of knocking out a gene for encoding a gamma chain of an interleukin-2 receptor in a mouse chromosome through a CRISPR/CAS9 technology, exons 1 and 2 of a gene of a gamma chain of an interleukin-2 receptor are selected as knockout sites, and Cas9 messenger RNA and guide RNA are jointly injected into fertilized eggs of a mouse through microinjection. The mouse with severe immunodeficiency cannot generate T, B and NK immune cells, so that the greatest convenient condition is created for transplantation of human hematopoietic stem cells.

The invention discloses a method for constructing a tumor xenograft model in mice immunized with gastric cancer based on an organ-like method, and an application thereof. The invention relates to a medical tumor animal model, based on an organ-like method. For the first time, the present invention employs a quasi-organ technique, that is, using a microcarrier as substrate. The composite human gastric cancer cell line SGC-7901/MKN-45 is transplanted subcutaneously into C57BL/6 mice to establish a tumor model of human gastric cancer with normal immune function. The tumor model constructed by this method has good tumorigenicity, and the tumorigenicity rate is up to 75%. The HE staining and the immunohistochemistry of the transplanted tumor accord with the characteristics of human gastric cancer. Compared with the existing immunodeficient mice transplanted tumor model, the model can better study and further clarify the mechanism of tumor occurrence and development in the normal immune system, and also provides a more valuable new animal model for the research and development of anticancer drugs. The invention is applied to the tumor model field.

The invention relates to the field of biomedicine. 5-fluorouracil (5-FU) is widely applied to the chemotherapy of gastral malignant tumors, but primary and acquired medicament resistant phenomena in the chemotherapy of colon cancer are universal; cell apoptosis is an important approach that chemotherapeutics play a role, and APAF-1 serving as regulatory protein is mainly involved in signal transduction of a mitochondrion approach of apoptosis; miR-23a can be combined with APAF1 serving as a target gene thereof so as to reduce the expression of the APAF1, and the apoptosis caused by the mitochondrion approach due to the chemotherapy is reduced so as to enhance the tolerance of the tumors to the chemotherapeutics; and the number of apoptotic cells is increased by reducing the expression amount of the in-vivo miR-23a so as to enhance the sensibility of the chemotherapeutics. The invention provides a PLKO-anti-miR-23a vaccine adjuvant for enhancing the chemotherapeutic effect of the chemotherapeutics, particularly the 5-FU. In-vitro experiments prove that the vaccine adjuvant can enhance the apoptosis level of colon cancer cells caused by the 5-FU serving as the chemotherapeutics obviously, and in-vivo experiments show that the vaccine adjuvant can enhance the chemotherapeutic effect of the 5-FU obviously, inhibit tumor growth and prolong the survival period of immunodeficient mice.

The invention relates to somatostatin gene defective immunodeficient mice, a preparation method and application thereof, in particular to a method for building somatostatin gene knocked-out SST-PNDG somatostatin combined immunodeficient mice on NOD-Prkdc and IL2rg-/-immunodeficient mice (PNDG mice), SST-PNDG somatostatin immunodeficient mice obtained thereby and application thereof. The mice can be used for biomedicine, pharmacy and clinical researches like heterograft, tumor growth and drug screening.

The invention relates to the field of medicines, relates to a method for establishing an animal model, and particularly relates to a method for establishing a human colon cancer transplanted carcinomamouse model. A preclinical tumor model is made through preprocessing mice with prostaglandin E2 and inoculating immunodeficient mice (Balb/c or NOD/SCID) with the in-vitro in-situ tumors of a patientagainst the defects of low establishing success rate and instability of existing colon cancer models. The invention provides a method for improving the success rate of a colon cancer PDX model, and especially provides the method for preparing the high-infiltration and high-metastasis stable human colon cancer transplanted carcinoma mouse model. The model has great values for preclinical evaluation, treatment and prognosis of the colon cancer.

An object of the present invention is to provide a mouse that enables the functions of human NK cell to be studied. A DNA consisting of a nucleotide sequence represented by SEQ ID NO: 1, which is a gene region comprising a DNA in which a cDNA sequence encoding interleukin 15 (IL-15) is operably ligated to a cDNA sequence encoding the signal peptide of human interleukin (IL-2), is inserted to immunodeficient mouse cDNA. In NOD-scid, IL-2rγ^null-hIL-15 Tg mice thus generated, hCD56⁺ cell having a concentration sufficient for conducting in vivo study on human mature NK cell are detected for at least 6 months after transplantation.

The invention belongs to the field of medicine, and relates to a method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on an organ-likemethod, and an application thereof. The aim of the invention is to use the organ-like culture technology, adopts microcarrier as scaffold material, and combines the liver cancer cells from patient toculture the cells. The invention also discloses an organ-like method for preparing the liver cancer cells-3D material complex, and then directly into that cell-3D material complex is inoculated subcutaneously into normal immunized mice to construct liver cancer transplanted tumor model from patients in vivo, which solved the problems of using nude mice, SCID mice and other immunodeficient mice, which are difficult to feed, expensive and lower tumor formation rate. Especially, it can solve the problem that human hepatocellular carcinoma transplanted tumor model in immunodeficiency mice can notreflect the important role of immune system in tumor occurrence and development, and human tumor transplanted tumor model in immunodeficiency mice has a long tumor formation cycle, which urgently needs the drug-sensitive demand of patients with clinical urgent need. The invention is applied to the field of mouse model construction.