782results about "Vascular endothelial cells" patented technology

A device for modelling physiological and pathophysiological states of the brain is provided. The device comprises a hydrogel with micropattern channels having a length, width and depth. The micropattern channels comprise cells selected from the group consisting of neurons, astrocytes, microglia, oligodendrocytes, neuronal organoids, cancer cells, cancer spheroids, brain tumoral cells and combinations thereof.

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 discloses a pharmaceutical composition for promoting angiogenesis and tissue regeneration and application thereof. The pharmaceutical composition of the present invention comprises an acidic secretory protein rich in cysteine, a disaccharide chain proteoglycan, and an XV-type collagen alpha1 chain. The biological material system loaded with the three types of proteins can promote angiogenesis, and can improve the tissue regeneration efficiency of bone, cartilage and skin defect areas.

A method for constructing a microfluidic bone organoid-on-chip can continuously observe the impacts of different external factors on a bone tissue structure based on the “ternary regulation theory” of bone angiogenesis-bone resorption-osteogenesis coupling. The method includes: (1) building a visual three-dimensional cell cultivation platform to continuously observe impacts of different external factors on a ternary regulation theory based on angiogenesis of osteoclast precursors, osteoblasts and osteoclasts, such that a user is allowed to directly observe interactions among various kinds of cells; (2) designing microfluidic channels in a chip to allow a reagent involved to present a concentration gradient distribution in the chip; and (3) adding three bioinks including osteoblasts, osteoclasts and vascular endothelial cells respectively, so as to reflect the three cell components individually, in pairs, and as a whole in the chip. An apparatus for constructing a microfluidic bone organoid-on-chip is further provided.

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 provides a construction method and application of a blood sinus networked liver organ. According to the method, pluripotent stem cells are co-differentiated into HAND1 < + > mesoderm and endoderm, a front intestine rear endoderm containing KDR < + > mesoderm progenitor cells is generated, a liver endoderm with CD34 < + > / CD31 < + > endothelial progenitor cells is generated, and the blood sinus networked liver organ is formed. The liver organ has a functional blood sinus endothelial network penetrating through the liver cluster, and has the characteristic functions of low-density lipoprotein uptake, human serum albumin removal, mediated secretion of blood coagulation factor 8 and the like. The method provides a new-generation highly bionic research tool for liver disease mechanism analysis, drug screening and regeneration repair.

In various aspects, the invention provides culture platforms, cell culture media, and methods of differentiating pluripotent cells into hematopoietic cells. In certain aspects, the invention also provides pluripotent stem cell-derived hematopoietic cells generated using the media and methods disclosed herein. The pluripotent stem cell-derived permanent HE cells produced by the methods provided herein are capable of differentiating into hematopoietic lineage cells comprising T cell progenitor cells and T cells in addition to NK cell progenitor cells, NK cells, NKT cells, or B cells.

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 discloses a construction method of induced pluripotent stem cells based on NFAT1 and application of the induced pluripotent stem cells in smoke diseases. The invention provides a construction method of induced pluripotent stem cells for preparing a smoke disease model, which comprises the following steps of: reprogramming CD34 + cells in in-vitro human PBMCs (OCT4 gene, SOX2 gene, KLF4 gene, c-MYC gene and NFAT1 gene are jointly introduced into the CD34 + cells); the obtained induced pluripotent stem cells are the induced pluripotent stem cells for preparing the smoke disease model. The invention constructs a pluripotent stem cell model capable of accurately simulating the pathological characteristics of smoke disease by introducing NFAT1. The model provides a brand new technical means for pathogenesis research, drug screening and personalized treatment of smoke diseases, and has important application value in the fields of disease model construction, tissue repair, precision medical treatment and the like.

The invention belongs to the technical field of organ chips, and particularly discloses a microvascular organ chip and a microvascular network culture method.The microvascular organ chip comprises a chip body, the chip body is internally provided with a microvascular culture channel and four culture medium channels, and the microvascular culture channel comprises a central culture cavity; the four culture medium channels are circumferentially arrayed around the central culture cavity and are respectively communicated with the central culture cavity, each of the four culture medium channels is provided with a first medium inlet and outlet hole, and the n culture medium channels are used as inflow channels of a culture medium; the 4-n culture medium channels are used as outflow channels of microvascular network outflow liquid in the microvascular culture channel, the inflow channels and the outflow channels form pressure differences to control interstitial flow of the microvascular network so as to promote complicated growth of the microvascular network, and n is larger than or equal to 1 and smaller than 4. Through the microvascular organ chip provided by the invention, a high-complexity and high-authenticity microvascular network can be cultivated.

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.

Disclosed is a method for producing T cells, the method comprising: (1) culturing three-dimensional cell aggregate(s) comprising cells that can differentiate into T cells, and stromal cells that express Notch ligand(s) derived from pluripotent stem cells. Also disclosed are T cells obtained by the method, and a medicine comprising the T cells.

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 present invention relates to a novel identified stem cell vMSC having angiogenesis characteristics, the vMSCs identified after being cultured under the culture conditions of the present invention are different from existing MSCs, are novel MSCs expressing CD141 cell surface antigens, have excellent cell expansibility, have an effect of directly forming blood vessels, have a paracrine effect of other MSCs, and are different from existing stem cells. The stem cell therapeutic agent for revascularization has a higher survival rate and a higher residual rate in tissues than conventional stem cell therapeutic agents, and can be used as a stem cell therapeutic agent for revascularization in which blood vessels are directly formed.

Platelet lysate compositions and cell culture media compositions for maintaining and / or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and / or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and / or flasks pretreated with the cell culture media compositions.

The present invention relates to a method for preparing an in vitro disease simulation model in which adult acute myocardial infarction and heart failure diseases are induced by ischemia-reperfusion injury using human pluripotent stem cell-derived cardiac organoids. The method includes: a step A of culturing human pluripotent stem cells to prepare a human pluripotent stem cell-derived cardiac organoid self-organized to include cardiomyocytes, fibroblasts, and vascular endothelial cells; a step B of culturing the human pluripotent stem cell-derived cardiac organoid prepared through step A in RPMI 1640 medium with cobalt (II) chloride added and glucose excluded to simulate ischemic conditions; a step C of culturing the human pluripotent stem cell-derived cardiac organoid that underwent ischemic condition simulation in step B in RPMI 1640 medium with D-glucose and calcium chloride added to simulate a reperfusion injury condition; and a step D of culturing the human pluripotent stem cell-derived cardiac organoid that underwent ischemia-reperfusion injury simulation in step C in RPMI 1640 medium with TGF-β and B-27 Supplement added to simulate a state of fibrosis resulting from heart failure.

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 provides a co-culture system of human alveolar epithelial cells and endothelial cells, which is used for researching interaction in an epithelial-endothelial barrier in lungs. It is indicated that infection with the highly pathogenic influenza A virus A / HK / 483 / 97 (H5N1) can significantly reduce alveolar fluid clearance (AFC) and increase protein permeability (APP). Administration of therapeutic extracellular vesicles (EV) after infection effectively restores AFC and APP, reduces viral replication, and enhances expression of antiviral genes. According to the invention, the treatment potential of EV is highlighted, and a valuable tool is provided for researching acute lung injury and improving a lung drug delivery technology.

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 application of a tumor-derived acellular extracellular matrix in preparation of a medicine for promoting angiogenesis and tissue regeneration. The invention provides a strategy for regulating and controlling tissue regeneration based on a tumor-derived acellular extracellular matrix microenvironment, a novel angiogenesis promoting biological material is constructed by combining the high similarity of a tumor microenvironment and a regeneration microenvironment such as angiogenesis, and high-efficiency regeneration is realized in various tissue defects. In addition, due to the characteristic of rapid proliferation of the tumor tissue, the productivity bottleneck of a traditional acellular extracellular matrix raw material source is broken through, and sustainable biological raw materials are provided for large-scale production. The method successfully solves the problem that a specific tissue-derived acellular extracellular matrix is difficult to regenerate in a complex damaged microenvironment.