544results about "Vascular endothelial cells" patented technology

The invention discloses a novel application of an RNF6 gene, namely, an application of screening a medicine for preventing and / or treating Zika virus infection, aiming at inhibiting expression of the RNF6 gene or knocking out the RNF6 gene. A series of functional experiments prove that the RNF6 gene promotes the replication of the Zika virus, and the purpose of reducing the replication of the Zika virus can be achieved by using an expression inhibitor of the RNF6 gene or knocking out the RNF6 gene; experimental results show that the RNF6 gene can be used as a potential target for preventing and / or treating Zika virus infection, and a new direction is provided for development of drugs for preventing and / or treating Zika virus infection targeting the RNF6 gene in the future.

The invention relates to the technical field of tumor biomedical engineering and organ chips, in particular to a vascularized gastric cancer organ chip and a preparation method thereof.The chip is composed of an integrated micro-fluidic main body, an annular micro-column array and an optical sealing film, and a central culture cavity is divided into a tumor area and a blood vessel area by micro-columns; the preparation method comprises the following steps: injecting a fibrous protein solution containing human umbilical vein endothelial cells and cancer-related fibroblasts into a vascular region, and adding thrombin for in-situ gelation to form a pre-vascularized network; the method comprises the following steps: mixing a patient-derived gastric cancer organ with a methacrylic acid esterified gelatin pre-polymerized solution, injecting the mixture into a tumor area, and carrying out photo-crosslinking immobilization; culturing for 7-14 days under the dynamic perfusion condition of a mixed culture medium to obtain a three-dimensional gastric cancer organ model containing a capillary network; the method is expected to be used for rapid, low-cost and high-throughput screening of chemotherapy or anti-angiogenesis drugs and accurate prediction of individual curative effects of patients.

The invention discloses a filling and repairing agent for promoting angiogenesis and a preparation method thereof, the filling and repairing agent is composed of a vascular endothelial cell supernatant, acellular matrix particles, a suspending aid and a protective agent, the vascular endothelial cell supernatant is prepared by inducing vascular endothelial cells into a tube under a low-oxygen three-dimensional condition, collecting a cell culture solution, centrifuging and concentrating; the acellular matrix particles are obtained by acellular tissue matrix, shearing, freeze-drying, liquid nitrogen grinding and collecting; according to the filling repairing agent for promoting angiogenesis, the vascular endothelial cell supernate and the acellular matrix particles are combined, the filling repairing agent contains high-concentration angiogenesis-related factors and a stable three-dimensional bionic scaffold structure, function complementation is achieved through the synergistic effect of the bioactive factors and the bionic scaffold, and the filling repairing agent for promoting angiogenesis is obtained. Therefore, the filling repair agent which can promote angiogenesis, construct a physical support framework and promote tissue repair and regeneration capacity is innovatively obtained.

The invention relates to a cell collection method capable of simultaneously collecting three cells in a co-culture model, and belongs to the technical field of biology. The invention provides a cell collection method capable of simultaneously collecting three cells in a co-culture model, and the cell collection method comprises the following steps: after a three-cell co-culture model is constructed, taking out a Transwell chamber, retaining cells in the lower chamber, and collecting the cells in the lower chamber; respectively digesting the cells on the two sides of the Transwell cell membrane by using a trypsin solution with the concentration of 0.5 g / 100mL so as to respectively collect the cells on the two sides of the Transwell cell membrane. According to the cell collection method disclosed by the invention, the three cells in the three-cell co-culture model are simultaneously collected in a manner of digesting the cells on the two sides of the Transwell membrane step by step by using pancreatin, so that not only is the cell and consumable cost saved, but also the experimental synchronism of the three cells is ensured, and convenience is provided for optimizing the experimental process.

The invention relates to a blood flow connection type multi-organ chip capable of being freely assembled and reused, the blood flow connection type multi-organ chip comprises a chip cover body, organ culture chambers, a chip matrix and a pressure-sensitive film, the chip matrix comprises a plurality of groups of organ culture units which are mutually independent, each group of organ culture units comprises at least two culture flow channels and a plurality of organ culture holes, and the culture flow channels are communicated with the pressure-sensitive film. The organ culture holes are sequentially distributed at intervals along the culture flow channel, the bottom end of each organ culture hole is communicated with the culture flow channel, each organ culture hole is used for accommodating an organ culture chamber, the culture flow channel is located at the bottom of the chip substrate, one end of the culture flow channel is communicated with the storage pool, and the other end of the culture flow channel is communicated with the connecting port; the chip cover body is arranged at the top of the chip base body and completely covers the storage pools and the organ culture holes, the pressure-sensitive film covers the bottom of the chip base body to seal the culture flow channels, and in each group of organ culture units, a circulation path is formed between at least two culture flow channels through the storage pools. The biological function of the human body is better simulated, and disease modeling and treatment strategy research are carried out.

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 bone organ chip loaded with a microelectrode array and a use method, and relates to the technical field of organ chips, the bone organ chip is made of a polydimethylsiloxane material, and the bone organ chip comprises a three-layer structure: a top layer, a middle layer and a bottom layer; the middle layer comprises two blood vessel micro-channels, two osteoblast micro-channels and a neuron micro-channel, the channels are separated through columns and micro-columns, and cell interaction in a bone micro-environment is simulated; a microelectrode array is arranged on the top layer and is used for monitoring electrical activity of neurons in real time; the bottom layer is provided with a pneumatic valve which can control opening and closing of the micro-channel. According to the invention, nerve and blood vessel channels are introduced, a nerve-blood vessel-immunoregulation bone microenvironment is constructed, physiological processes such as osteogenesis-blood vessel formation coupling and the like are embodied, intervals of different sizes can enhance intercellular crosstalk and allow blood vessels and neuron axons to selectively pass through, so that the chip can simulate a complex microenvironment of bone tissues, and the microenvironment of the bone tissues can be simulated; and an efficient and accurate in-vitro model is provided for bone disease mechanism research and drug development.

The invention discloses a preparation method and application of a novel hybrid extracellular vesicle. According to the invention, vesicles derived from endothelial cells and neutrophil cells are combined with deferoxamine, and a biological mixed nano vesicle platform (DFO (HEVS)) is developed to solve the problem of diabetes wound healing. According to the dual-targeting system, DFO is accurately delivered to a wound part by utilizing CXCR4 mediated endothelial cell homing and beta2 integrin dependent inflammation tropism, DFO (at) HEVs activates and recovers vascular regeneration through HIF-1alpha / VEGF, ferroptosis is inhibited through Nrf2 / GPX4 signal transduction, macrophages are reprogrammed into a repair promoting M2 phenotype, and oxidative stress-inflammation-ferroptosis circulation is effectively broken.

The invention relates to a retina nerve and blood vessel unit in-vitro three-culture model and a preparation method thereof, and belongs to the technical field of biology. The invention provides an in-vitro three-culture model of a retina nerve and blood vessel unit. The in-vitro three-culture model is obtained by carrying out in-vitro Transwell indirect co-culture on retina microvascular endothelial cells, retina ganglion cells and retina Muller cells. The in-vitro three-culture model of the retina nerve and blood vessel unit is based on a three-cell indirect co-culture method, retina microvascular endothelial cells, retina ganglion cells and retina Muller cells are simultaneously incorporated by utilizing Transwell, and the three cells are in indirect contact. The in-vitro three-culture model of the retina nerve and blood vessel unit can be used for more objectively simulating pathophysiological states of various cells of the retina nerve and blood vessel unit, so that common pathophysiological changes of mutual dependence and interaction of the three cells under in-vitro conditions can be observed.

The invention provides universally acceptable “off-the-shelf” hypoimmunogenic pluripotent cells and differentiated cardiac, endothelial, neuronal, islet, or retinal pigment cells thereof. Such hypoimmune cells are used to treat patients in need thereof. The cells lack major immune antigens that trigger immune responses and are engineered to avoid phagocytic endocytosis.

The invention provides a vascularized bone organ and a construction method thereof, and a vascularized bone organ model containing a hollow vascular network is constructed by combining a suspension 3D biological printing method, a cell-carrying printing strategy and a sacrificial biological ink material. The vascularized bone organ model has excellent structural stability, mechanical performance, growth and development capability and osteogenesis capability, selected materials and a manufacturing method are simple in process, engineering stable production can be achieved, cells can be blended for a long time for three-dimensional culture, and the vascularized bone organ model has high applicability and research potential.

The invention discloses application of nano-iron in vascular endothelial cells, and belongs to the technical field of cell engineering and genetic engineering. According to the invention, PEI coated Fe3O4 is taken as a research object, the application of PEI coated Fe3O4 in vascular endothelial cells is researched by adopting a molecular and cell biology method, and regulation of PEI coated Fe3O4 on vascular endothelial regeneration of the vascular endothelial cells after myocardial infarction is verified from multiple levels and multiple angles. The invention can provide a new reference for the recovery of cardiac functions after myocardial infarction, and has good application value.

Disclosed are methods for preparing non-filamentous scaffolds (e.g., sheets) for cell or tissue culture. These methods can comprise providing at least a first printing composition (e.g., a bioink) and a second printing composition (e.g., a bioink or a fugitive ink); chaotic printing the first printing composition and the second printing composition to generate a microstructured precursor comprising a plurality of lamellar structures formed from the first printing composition and the second printing composition; extruding the microstructured precursor through a nozzle (e.g., a fan-shaped nozzle, a curved fan-shaped nozzle, or an annular nozzle) to produce a non-filamentous microstructured precursor; and curing the non-filamentous microstructured precursor to provide the non-filamentous scaffold for cell or tissue culture.

Methods for generating human vascular leptomeningeal cells (VLMCs) from human oligodendrocyte progenitor cells (OPCs) are provided using chemically-defined culture media that allow for generation of differentiated cells in a 34-day culture protocol. Methods of generating human VLMCs from human pluripotent stem cells using chemically-defined culture media in a 40 day culture protocol are also provided. Culture media, isolated cell populations, and kits are also provided.

The invention discloses a preparation method of a cochlear organ chip integrated with a blood labyrinth barrier, and belongs to the field of biomedical engineering. The preparation method of the simulated blood labyrinth barrier comprises the following steps: constructing an endothelial cell culture cavity and a pericyte culture cavity, and separating the endothelial cell culture cavity and the pericyte culture cavity through a simulated basement membrane; and respectively inoculating percutaneous cells and endothelial cells into a percutaneous cell culture cavity and an endothelial cell culture cavity, and culturing to obtain the simulated blood lost barrier. The chip can effectively simulate the structure and the function of the BLB, and the barrier integrity of the BLB is evaluated through TEER (trans-epithelial electrical resistance) measurement and an apparent permeability coefficient. The invention also relates to application of the chip in NIHL drug screening, in particular to drug evaluation for oxidative stress induced inner ear organ injury. Through the platform, the protection effect of the candidate drugs on the TBHP-induced oxidative stress injury of the inner ear organs can be evaluated, and a new tool and thought are provided for developing novel NIHL treatment drugs.

The invention discloses a 3D (three-dimensional) visual bionic intestine-blood vessel-nerve interface organ simulation chip, which is characterized in that a micro-fluidic chip is used as a platform to construct an intestinal tract-blood vessel-brain system which simulates biological transformation of perfluorinated compounds in intestinal tracts under near physiological conditions and biologically transports the perfluorinated compounds through blood vessels; a neuronal cell culture chamber, a vascular endothelial cell culture chamber and an intestinal tract cell culture chamber are respectively arranged on the micro-fluidic chip, the culture chambers are communicated through micro-columns, and the vascular endothelial cell culture chamber serves as a barrier in the middle; perfluorinated compounds enter corresponding cell culture cavities for culture through reagent injection channels of different cell culture channels, generated metabolites are discharged through the channels, and multi-organ interaction and toxicity induced by the perfluorinated compounds are analyzed. The chip can simulate an intestinal barrier structure, a blood vessel and a neural network structure, can highly simulate biological transport and transformation of a perfluorinated compound between intestines and brains, and effectively reduces cost and time.

The invention discloses a brain glioma organ chip with adjustable tumor microenvironment gradient and a method for mediating formation of brain glioma organoid by the brain glioma organ chip, and belongs to the technical field of organ chips. The preparation method comprises the following steps: firstly, preparing a glioma organ chip with adjustable tumor microenvironment gradient by utilizing a prepolymer solution containing glioma cells and vascular endothelial cells and a prepolymer solution containing biochemical molecules; then, the obtained brain glioma organ chip with the adjustable tumor microenvironment gradient is subjected to three-dimensional dynamic co-culture under the perfusion condition, three-dimensional culture and co-culture of brain glioma cells and vascular endothelial cells are achieved at the same time, endothelial-glioma multicellular tumor spheres are formed, and construction of brain glioma organs is achieved. The brain glioma organoid established by the invention successfully summarizes key characteristics of a tumor microenvironment, including biophysical factors simulated by physiological fluid and biochemical factors of three-dimensional culture microgel with adjustable concentration gradient of biochemical molecules.

The invention provides recombinant III-type collagen and application thereof, and relates to the technical field of genetic engineering. According to the invention, the yield of the collagen is improved by optimizing the nucleotide sequence of the target gene and optimizing the culture conditions of pichia pastoris, so that the collagen is efficiently expressed in engineering bacteria. The recombinant engineering bacterium shows strong potential in the aspect of improving the yield of protein with high economic value, and is suitable for large-scale industrial production. The recombinant III-type collagen is high in expression quantity, simple in preparation process and suitable for preparation of biological materials for medical cosmetology, wound repair and the like, and has a wide application prospect.

The embodiment of the invention discloses a method for constructing a cartilage micro-tissue by compounding an extracellular matrix with a vascular matrix component and application. The method comprises the following steps: preparing a porcine decellularized cartilage extracellular matrix; generating a cartilage extracellular matrix suspension; carrying out screening treatment on the cartilage extracellular matrix suspension; preparing a porcine acellular cartilage extracellular matrix microcarrier; preparing a target suspension of a common system from the porcine acellular cartilage extracellular matrix microcarrier and vascular matrix component cells, and transferring the target suspension to a bionic floating microenvironment of a shaker bioreactor for dynamic culture; carrying out culture treatment on the porcine acellular cartilage extracellular matrix microcarrier; and taking out the cell-loaded porcine decellularized cartilage extracellular matrix microcarrier from the cultured and treated porcine decellularized cartilage extracellular matrix microcarrier as the constructed cartilage micro-tissue. According to the embodiment, the condition of secondary injury to a user can be improved and reduced, the repairing effect is improved, and the adaptability of the constructed cartilage micro-tissue is improved.

The invention belongs to the technical field of organ chips, provides an in-vitro brain lymphatic system chip construction method and a brain lymphatic system chip, and makes up the blank of brain lymphatic system in-vitro model construction. The method comprises the following steps: firstly, carrying out flushing treatment, disinfection and sterilization treatment, rinsing treatment and cell culture on a micro-fluidic channel in the chip; then, in the treated micro-fluidic channel, blood vessel channel cell inoculation and brain parenchyma channel cell inoculation are carried out; and finally, in the blood vessel channel, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, basic formation of in-vitro cerebral vessels is achieved after continuous perfusion, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, and in-vitro cerebral vessels are formed. Intermittent liquid flow simulating arterial fluctuation is achieved.

The invention relates to a phenylacetylglutamine / phenylacetylglycine induced vascular smooth muscle cell senescence model and an establishment method and application thereof, and belongs to the technical field of cell models.The method comprises the following steps that vascular smooth muscle cell suspension is placed in a porous plate, 10% of FBS and 1% of mycillin are added into a culture medium, 24 h after cells adhere to the wall, the vascular smooth muscle cell senescence model is obtained, and the vascular smooth muscle cell senescence model is obtained. The method comprises the following steps of: adding a proper amount of phenylacetyl glutamine or phenylacetyl glycine dimethyl sulfoxide solution into a porous plate, changing a serum-free culture medium, continuously culturing for 24 hours, adding a proper amount of phenylacetyl glutamine or phenylacetyl glycine dimethyl sulfoxide solution into the porous plate, changing the culture medium added with phenylacetyl glutamine or phenylacetyl glycine once every 24 hours, and collecting cells after 72 hours. According to the invention, phenylacetylglutamine / phenylacetylglycine is used for inducing and establishing the vascular smooth muscle cell senescence model for the first time, the method is easy to operate and low in cost, and a foundation is laid for researching the relationship between cardiovascular diseases and senescence.

The present invention provides methods of constructing vascularized retinal organoids by differentiating retinal progenitor cells and endothelial cells. The method is co-cultured by differentiating retinal progenitor cells and endothelial cells, and comprises the following steps: carrying out multiplication culture on pluripotent stem cells until the cell confluence degree is 80-90%, and digesting; carrying out differentiation culture on the digested pluripotent stem cells in a culture medium containing retina induction factors to obtain retina progenitor cells; and co-culturing the retinal progenitor cells differentiated for 12-13 days and endothelial cells to obtain the vascularized retinal organ, wherein the endothelial cells are obtained by any one method of induced differentiation of digested pluripotent stem cells or culture of endothelial cell lines. The vascularized retinal organ with the retinal layered structure is constructed by integrating the endothelial cells and the retinal progenitor cells for the first time, and the limitation that in the prior art, the retinal organ lacks vascularization is successfully overcome.

Several embodiments relate to methods of repairing and / or regenerating damaged or diseased tissue comprising administering to the damaged or diseased tissues compositions comprising exosomes. In several embodiments, the exosomes comprise one or more microRNA that result in alterations in gene or protein expression, which in turn result in improved cell or tissue viability and / or function.

The present disclosure relates to organoids and particularly to brain organoid models. The brain organoids include neurons, microglia, astrocytes, and blood vessels. The brain organoid models can be used to create models for Alzheimer's Disease. Methods of using the brain organoids for drug discovery are also described.

The invention relates to the technical field of cell culture, in particular to an oriented porous bionic gelatin scaffold for controlling cell orientation arrangement, a preparation method and application thereof, and the oriented porous bionic gelatin scaffold can be used for cell culture meat production. The preparation method comprises the following steps: mixing and solidifying gelatin and glutamine transaminase, directionally freezing, freeze-drying, sterilizing, inoculating and culturing cells and the like. Parameters and culture medium components in each step are also explained. Compared with the prior art, a regular directional porous structure is formed through a hydrogel scaffold and directional freezing, directional arrangement and growth of fish muscle satellite cells are guided, the cell activity is high, and the proliferation and differentiation effects are good; the gelatin material is adopted, so that the cost is low, the biocompatibility is good, and the gelatin material is superior to collagen and wheat glutelin; the preparation process is simple, does not need complex equipment, is suitable for large-scale production, and promotes the development of cell culture meat.

Methods are provided for the step-wise generation of endothelial cells. Also provided are methods for generating pure populations of such cell types and derivatives thereof. The instant disclosure also provides methods of screening for cellular responses of the generated cell types and derivatives thereof. Treatment methods making use of the generated cell types and derivatives thereof are also provided. The instant disclosure also provides systems, compositions, and kits for practicing the methods of the disclosure.

A heart organoid of at least 60% cardiac cells irrespective of optional additional cells of a vascular system of the organoid, or including at least 50% cardiac cells; wherein the cardiac cells surround an inner cavity, wherein the cardiac cells are selected from the group of cardiomyocytes, endocardial cells and epicardial cells, and wherein the heart organoid includes cardiomyocytes or endocardial cells that directly face the inner cavity, a method for the generation of such a tissue model and uses thereof.

The invention belongs to the technical field of regenerative medicine, and particularly relates to a method for generating vascular endothelial cells by inducing pluripotent stem cells to differentiate in vitro. The method comprises the following steps: carrying out first culture on pluripotent stem cells in a differential medium I to obtain cells A; carrying out second culture on the cell A in a differential culture medium II, and differentiating and directionally forming an endoderm cell B; performing third culture on the endoderm cells B in a differential culture medium III to obtain differentiated cells C; carrying out fourth culture on the differentiated cells C in a differentiation culture medium IV, and differentiating to form differentiated cells D; carrying out fifth culture on the differentiated cells D in a differential culture medium V, and differentiating to obtain vascular endothelial cells; wherein the differential culture media I-V do not contain activin A, VEGF (vascular endothelial growth factor) and BMP4 (bone morphogenetic protein 4). The method can improve the differentiation efficiency of the IPS cells to the vascular endothelial cells, and has a wide prospect in the cell therapy industry.

The invention relates to the related technical field of molecular biology and cell engineering, in particular to an FLP-IN-PIEC cell line and a construction method and application thereof, and the construction method comprises the following steps: step 1, constructing CRISPR / Cas9 site-specific cleavage plasmid containing a pig HIPP11 safety site sequence; step 2, constructing a donor plasmid Donor-FRT containing a homologous arm of a safety site of the HIPP11; 3, jointly transfecting plasmids in the step 1 and the step 2 into a porcine hip artery endothelial cell line (PIEC) to obtain an FLP-IN-PIEC cell line for expressing red fluorescence; and 4, under the action of Flp recombinase, transfecting an exogenous gene donor plasmid containing an FRT site to the FLP-IN-PIEC cell line obtained in the step 3, and driving the expression of a target gene through an optimized strong promoter to realize fixed-point transfer and efficient expression of the exogenous gene in the pig (PIEC) cell line. Different from an existing Flp-In system depending on a random integration strategy, the construction method disclosed by the invention emphasizes fixed-point integration and promoter optimization, and improves the homogeneity, stability and screening efficiency of exogenous gene expression.

Methods for producing prevascularized organoids, and compositions comprising such prevascularized organoids, are provided. Methods of treating ischemic conditions and promoting angiogenesis by administering the prevascularized organoids to a subject in need thereof are also provided.