3656results about "Artificial cell constructs" patented technology

The invention provides a pourable tumor organoid vascularization model and a construction method thereof, and relates to the technical field of organoid culture. A temperature-sensitive degradable material and a photo-crosslinking modified temperature-sensitive material are used, the composite stent is printed through two channels, a multi-component gel mask partition curing process is combined, the stability of a perfusion pipeline is improved, and a partitioned highly-bionic complex microenvironment is accurately constructed in a macroscopic three-dimensional space; and constructing to obtain the perfusion tumor organ vascularization model. The model is high in bionic degree and stable in structure, can be applied to deepening cell and microenvironment interaction research and tumor-related drug screening, and has a good application prospect.

The invention discloses a 3D intestinal organ differentiation method based on human pluripotent stem cells and an induction culture medium and application thereof, and relates to the technical field of stem cells. According to a culture medium formula combination, intestinal organs can be differentiated into various cell types such as epithelial cells, neuroendocrine cells and endothelial cells; the method is a key mark for successful differentiation and functional maturation of intestinal organs. According to the 3D intestinal organ differentiation method disclosed by the invention, histological structures such as intestinal crypts are differentiated from intestinal organs generated by differentiation, and the intestinal organs can creep in a maintenance stage, so that the intestinal organs are changed from structural bionics to functional simulation, and the significance of the 3D intestinal organ differentiation method is far better than that of pure morphological simulation. Through a systematic culture medium formula, a clear operation process and a multi-stage induction strategy, the 3D intestinal organ with structural integrity, cell diversity and functional activity is successfully constructed, and the system provides an efficient, reliable and extensible in-vitro model platform for intestinal biological research and related application.

The invention relates to an organ-like core-shell microsphere as well as a preparation method and application thereof. The preparation method comprises the steps that 1, raw materials are prepared, specifically, shell raw materials and inner core raw materials are prepared, the shell raw materials comprise a photoinitiator and methacrylated hyaluronic acid (HAMA), and the inner core raw materials comprise organoid precursor cell suspension and matrigel; 2) molding: extruding the dispersion phase of the shell raw material wrapping the core raw material to a continuous phase by using a micro-fluidic chip and a high-precision injection pump, cutting the dispersion phase into liquid drops by the continuous phase, and performing illumination curing on the liquid drops to form core-shell microspheres; and (3) incubating, namely culturing the core-shell microspheres in a culture medium, so that the organoid precursor cells in the inner core are developed into organoids. According to the preparation method, rapid forming and curing of the organ-like precursor cells carried by the matrigel are achieved, so that the time of the organ-like precursor cells staying in the oil phase is shortened, the cells can obtain oxygen and nutrient substances, and the cell activity is improved.

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 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 provides a method for separating extracellular vesicles from a milk source, which comprises the step of adding a dispersing agent before adding a precipitator into the milk source to precipitate casein so as to reduce the wrapping of protein precipitation on the extracellular vesicles and improve the yield of the extracellular vesicles, thereby obtaining a milk source extract rich in the extracellular vesicles.

The invention belongs to the field of biological medicines, and particularly discloses application of a Ptprj agonist GJ103 in preparation of a medicine for treating acute kidney injury (AKI) induced by cisplatin. In-vivo and in-vitro experiments prove that the GJ103, by activating Ptprj, can significantly down-regulate expression of pro-apoptotic protein Bax and Cleved Caspase-3, up-regulate anti-apoptotic protein Bcl2 and reduce infiltration of inflammatory factors TNF-alpha and IL-6, so that apoptosis and inflammatory response of renal tubular epithelial cells are relieved. In in-vivo experiments, GJ103 (20-40mg / kg / day) can reduce serum creatinine and urea nitrogen levels of cis-platinum model mice and improve pathological injuries such as renal tubule dilatation; in in-vitro experiments, 20-40 [mu] M of GJ103 can inhibit apoptosis of renal tubular epithelial cells and reduce expression of renal injury markers NGAL and Kim-1. The pharmaceutical composition contains GJ103 and a pharmaceutical carrier, the preparation form can be a 4mg / mL injection (the purity is greater than or equal to 99.46%) or an oral preparation, and a new strategy is provided for clinical treatment of cisplatin renal toxicity.

The invention provides an in-vitro skin blood vessel immune model as well as a preparation method and application thereof. THP-1 human monocyte leukemia cells are induced to be differentiated into M0 type macrophages, and then the M0 type macrophages are respectively induced into M1 type macrophages and / or M2 type macrophages; then co-culturing the M1 type and / or M2 type macrophages and vascular endothelial cells to form a vascular immune model; finally, the 3D skin model is placed on the blood vessel immune model, external stimulation is conducted, and the in-vitro skin blood vessel immune model is constructed. The in-vitro skin blood vessel immune model can be used for repairing skin barriers, inflammation pathways, blood vessel metabolism, the expression level of genes or proteins related to extracellular matrixes, vascular endothelial cells, the proliferation and migration ability of activated macrophages and the like. The seven feature dimensions of the physiological structure feature of the skin model are used for screening the to-be-tested sample and exploring the action mechanism, and the method has the characteristics of rapidness, multiple screening dimensions, high accuracy, low construction difficulty, low cost and high universality.

The invention relates to a method for inhibiting in-vitro infection of a porcine epidemic diarrhea virus by using JPH203 and application. The JPH203 can be used for preparing an antiviral drug for inhibiting infection of the porcine epidemic diarrhea virus. JPH203 with the concentration of 5 [mu] M is added in the process that Vero cells are infected with PEDV, compared with a control group without JPH203, the virus infection condition, the protein expression quantity, the RNA content and the virus titer of the PEDV in a treatment group with JPH203 are all remarkably reduced, and it is indicated that JPH203 can be used for inhibiting in-vitro infection of the porcine epidemic diarrhea virus.

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 relates to the technical field of repair materials, and particularly discloses an extracellular matrix fiber material as well as a preparation method and application thereof. The invention discloses a preparation method of an extracellular matrix fiber material. The preparation method comprises the following steps: taking animal tissues, and carrying out virus inactivation, decellularization and antigen removal, degreasing, drying, grinding and sterilization; the decellularization and antigen removal method comprises the following specific steps: sequentially soaking a sample with a surfactant solution for h; then, soaking treatment is carried out by adopting a salt-alkali solution which contains 0.05-3 mol / L of salt and has the pH value of 8-11; then carrying out soaking treatment by adopting a 0.1-1wt% enzyme solution; and in degreasing, one or more of normal hexane, trichloromethane, dichloromethane, methanol, ethanol, acetone, diethyl ether and ethyl acetate are used for soaking the sample. According to the technical scheme, the ECM powder particle material with high growth factor content is prepared.

The invention provides a micro-fluidic chip capable of culturing oral squamous cell carcinoma organoid and a preparation method thereof, and relates to the technical field of micro-fluidic chips, the chip comprises a four-layer structure: the first layer is a closed layer, the second layer is a culture solution / drug pool layer (containing a concentration gradient injection channel and a slow flow channel), the third layer is a matrix culture pool layer, and the fourth layer is a matrix culture pool layer. The fourth layer is a matrigel inflow layer (containing an S-shaped flow channel); the second layer and the third layer are integrally formed, and all the layers are connected through oxygen plasma welding; the problems that in the prior art, matrigel is prone to shifting, drug gradient regulation and control is poor, operation is tedious, and adaptability is insufficient are solved, the matrigel and organoids can be stably fixed, the drug concentration gradient can be precisely regulated and controlled, the nutrition supply efficiency is enhanced, the structural integration level and operation convenience are improved, and the application prospect is wide. The method is suitable for oral squamous cell carcinoma organoid culture, drug screening and tumor mechanism research.

The invention discloses a single-shaft adjustable mechanical stretching device for organoids or cells and a stretching culture method. A fixed support and a guide rail are arranged on a fixed base through a guide rail lifting seat, and a sliding block above the guide rail is connected with a stretching movement end through an adapter seat; the stretching moving end and the stretching fixed end are arranged at an interval, a lead screw of a stepping motor on the fixed support is fixedly connected to a coupler, and the coupler is fixedly connected with an adapter through a blocking block; handles on the two sides of the culture chamber are fixed to a stretching fixing base and a stretching fixing end respectively, a stepping motor is started, a lead screw of the stepping motor drives an adapting base to move longitudinally when moving, the longitudinal movement of the adapting base drives a sliding block to move on a guide rail, and then a stretching movement end is made to be far away from / close to the corresponding stretching fixing end; according to the invention, accurate and controllable tensile mechanical stimulation is carried out on different forms of organoids or cells through the replaceable culture chamber and the accurately controllable stepping motor, so that the tissue maturity / calcification degree is obviously changed or the cell differentiation process is adjusted.

The invention provides a specific sequence nucleic acid molecule and application thereof as a PDRN, and relates to the technical field of biology. A novel nucleic acid molecule with a specific sequence is obtained, and the sequence has certainty, biological activity and stable product quality. The PDRN of the specific sequence does not have cytotoxicity, is high in safety and has a wide application prospect. Moreover, compared with the extracted PDRN, the PDRN with a specific sequence is found for the first time, the product has a remarkable effect of promoting VEGF expression, cell damage can be effectively repaired, and a novel skin-care and repairing functional component is provided for the industries of cosmetics and medicines. The invention further provides a preparation method of the PDRN with the specific sequence, and the preparation method is simple and suitable for popularization, application and industrial production.

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The invention provides a pancreatic islet organ capable of efficiently maintaining pancreatic islet beta cell identity and a preparation method and application thereof. Specifically, the invention provides the preparation method for the in-vitro pancreatic islet organ, and the preparation method comprises the following steps: carrying out induced differentiation culture on the ZnT8-knocked-out pluripotent stem cells, so as to obtain the mature pancreatic islet organ. The invention also provides a corresponding transplantation composition and application thereof. The invention discloses that ZnT8 knockout is a key factor for effectively maintaining the identity stability of beta cells in pancreatic islet organs. In addition, the pancreas islet beta cell identity maintenance agent is combined with the ZnT8-knocked-out pancreas islet organ for use, so that the identity stability of the pancreas islet organ beta cell can be obviously maintained. The invention provides a new strategy for transplantation treatment of type 2 diabetes, and has excellent clinical transformation value.

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 belongs to the technical field of biology, and particularly relates to a high-throughput preparation method of a single-cell-derived organoid based on a microcapsule spherical shell structure, and the preparation method comprises the following steps: preparing an aqueous two-phase microcapsule encapsulated with a single cell, culturing the single cell in the aqueous two-phase microcapsule to obtain the organoid, the substances of the inner phase and / or the outer phase respectively comprise any one or a combination of more of the following substances: methacrylated gelatin, carboxymethyl cellulose, glucan, sodium alginate and polyvinyl alcohol. The invention also provides the organoid prepared by the preparation method and application of the preparation method. According to the invention, the microcapsule with a permeable core-shell structure is formed by packaging the microcapsule in a single-cell form in two-aqueous-phase liquid drops, the wrapping rate is effectively controlled and adjusted on the premise of conforming to Poisson distribution by optimizing parameters, and the single-cell wrapping rate and the organoid single-cell induction efficiency are optimized.

Provided herein are methods and systems for improving performance of three-dimensional printing systems, which may include: printing a first portion of the 3D object using a first parameter set and a first light beam, wherein the first parameter set includes at least one first parameter corresponding to a first optical property of the first light beam; and printing a second portion of the 3D In object different from the first portion using a second parameter set and a second light beam, wherein the second parameter set includes at least one second parameter corresponding to a second optical property of the second light beam, wherein the second parameter set is different from the first parameter set, wherein the second optical property is different from the first optical property, to yield at least at least a portion of the 3D object comprising the first portion and the second portion.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

To provide novel transduction enhancers and uses thereof.SOLUTION: The present invention relates to a method for transducing a target cell, the method comprising the step of contacting a target cell with a retroviral vector and a compound capable of enhancing transduction efficiency or a combination of such compounds, wherein the target cell is pre- and / or co-stimulated by pre- and / or co-incubation with a transduction enhancing compound or a combination of transduction enhancing compounds prior to and / or during contacting the target cell with the retroviral vector.SELECTED DRAWING: None

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

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.

The invention relates to the technical field of polypeptides, in particular to an enzyme-mimetic polypeptide, hydrogel, a preparation method and application, the enzyme-mimetic polypeptide selects a peptide fragment I and a peptide fragment II with biocompatibility and degradability as basic sequences, and the peptide fragment I and the peptide fragment II form a beta folding unit, so that the stability of a polypeptide structure can be enhanced; then, the peptide chain is driven to be self-assembled to form a regular nanofiber structure; according to the invention, the characteristic that the biomimetic polypeptide and metal ions are subjected to a complex reaction to form a chelate is utilized, the transparent and stable hydrogel can be rapidly formed without adding a gelatinizing agent, and TEM, SEM and rheological characterization are carried out on the hydrogel to find that the hydrogel containing the biomimetic polypeptide has a porous nanofiber network structure, so that the biomimetic polypeptide can be used for preparing the hydrogel. Meanwhile, the biomimetic polypeptide hydrogel has the same flexibility as natural tissues, and the characteristics indicate that the biomimetic polypeptide hydrogel is expected to have wide application prospects in the aspects of biomimetic materials, antibiosis, cell culture, drug carriers and the like.

The invention relates to a Vero cell serum-free culture medium and application thereof, the serum-free culture medium is suitable for free suspension growth of Vero cells and is free of animal sources and protein components, adherent cells can be directly domesticated through a certain method to screen out Vero suspension cells, and serum-free suspension culture of the Vero suspension cells is achieved in the serum-free culture medium. The density of cells subjected to suspension culture by adopting the serum-free culture medium can reach more than 107 cells / mL, the cell viability is more than 90%, the cells are well dispersed, the cells can still normally grow and proliferate after continuous subculture for 20 generations, and the cells are sensitive to various viruses.

The invention belongs to the field of biotechnology and aquaculture, and particularly relates to a method for improving hypoxia resistance of pelteobagrus vachelli based on hypoxia induction of liver extracellular vesicles of the pelteobagrus vachelli. The method comprises the following steps: extracting high-purity extracellular vesicles EVs in liver tissues of pelteobagrus vachelli subjected to low-oxygen treatment by adopting a method of combining differential centrifugation and high-precision iodixanol density gradient centrifugation; diluting the extracted EVs to 2mu g / mu L by using PBS (Phosphate Buffer Solution) to prepare an EVs suspension; an intraperitoneal injection mode is adopted, the EVs suspension is injected into the body of the pelteobagrus vachelli, and after injection is conducted for 24 hours, the hypoxia resistance of the pelteobagrus vachelli is evaluated through a suffocation point detection test. The invention opens up a brand new path for improving the hypoxia resistance of the pelteobagrus vachelli, provides a new idea and a new method for efficiently enhancing the hypoxia resistance of economic fishes in the field of aquaculture, is expected to play an important role in actual aquaculture production, and assists the sustainable development of the aquaculture industry.

The invention discloses a mouse lung organ and fibroblast co-culture method, which comprises the following steps of: performing in-vitro mouse lung organ culture on mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, and after the lung organs in the Transwell upper chamber are differentiated, replacing with a lower chamber paved with mouse lung fibroblasts, therefore, a gas-liquid interface co-culture system of the mouse lung organ (upper chamber) and the fibroblast (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the upper chamber organ and the phenotype, function and related signal path activation level of the lower chamber fibroblast can be dynamically researched.