2036results about "Vertebrate cells" patented technology

The invention provides an artificial nucleic acid molecule which is used for improving the expression quantity of target amino acid, polypeptide or protein. The artificial nucleic acid molecule at least comprises a target 5'untranslated region (UTR), a target coding region (CDS) and a target 3 'untranslated region (UTR). Wherein the sequence of the target 5 'UTR is one of the following sequences: 5' UTR of a high-expression gene and a 5 'UTR variant of the high-expression gene. The sequence of the target 3 'UTR is one of the following sequences: 3' UTR of a high-expression gene and a 3 'UTR variant of the high-expression gene. Optionally, the artificial nucleic acid molecule may further comprise, for example, a 5 '-end cap structure (Cap), a PolyA tail. The 5 'UTR and the 3' UTR have regulating effects on translation and stability of nucleic acid molecules, so that the 5 'UTR, the 3' UTR and variants thereof are selected from high-expression genes, the nucleic acid molecules can be further stabilized and are not easy to degrade, and the amount of protein or polypeptide obtained by translation of the nucleic acid molecules can be increased. The invention also provides methods for making, delivering, and using such artificial nucleic acid molecules, as well as the use of the artificial nucleic acid molecules for the treatment and / or prevention of related diseases or disorders.

The invention discloses a novel application of a small molecule compound BRD4780 in resisting ricin, the small molecule compound BRD4780 has a remarkable effect of resisting ricin toxicity and can be used in development of anti-ricin drugs, a brand new candidate molecule is provided for development of the anti-ricin drugs, the drug research and development blank in the field is filled, and the application of the small molecule compound BRD4780 in resisting ricin is developed. The method has important significance in the aspects of clinical treatment, biological protection and immunotoxin treatment safety optimization, and is wide in application prospect.

A mammalian or avian cell line that expresses high levels of human influenza virus receptors is provided. In one embodiment, the cell line supports human influenza virus, e.g., human A / H3 influenza virus, isolation and growth much more effectively than corresponding conventional (unmodified) cells or in corresponding human virus receptor-overexpressing cells, and the propagated viruses may maintain higher genetic stability than in the corresponding cells.

The invention discloses an effect of Collagen in relieving vomitoxin injury of bovine rumen epithelial cells by vitamin E, and belongs to the technical field of genetic engineering, the Collagen is COL1A1 protein, COL2A1 protein and COL5A1 protein; the amino acid sequence of the COL1A1 protein is as shown in SEQ ID NO. 19; the amino acid sequence of the COL2A1 protein is as shown in SEQ ID NO. 20; the amino acid sequence of the COL5A1 protein is as shown in SEQ ID NO. 21. According to the effect of Collagen in relieving bovine rumen epithelial cells damaged by vomitoxin through vitamin E, it is found that vitamin E can effectively protect the cell membrane integrity of the bovine rumen epithelial cells and can relieve cell viability decline, oxidative stress, immune and inflammatory reactions and the cell apoptosis rate caused by DON; vitamin E can promote ECM receptor effect and protein digestion and absorption ways by adjusting Collagen protein so as to protect rumen epithelial cells of dairy cows.

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 present invention belongs to the field of virus detection technology, and specifically relates to monoclonal antibodies with neutralizing activity against adenovirus type 5 and their uses. The monoclonal antibody 8B9 of the present invention can specifically recognize and neutralize adenovirus type 5, and the amino acid sequences of the heavy and light chain variable regions and their complementarity determining regions of the monoclonal antibody 8B9 are provided. The present invention also provides an in vitro neutralization activity assessment model based on HEK-293A cells. At a concentration as low as 3.3 μg / mL, the titer of the neutralizing activity against the virus stock solution after 6000-fold dilution is 7×10 4 TCID 50 / mL of virus has close to 100% neutralizing activity. After further humanization, it is expected to be developed into a neutralizing antibody drug for the treatment of diseases such as severe pneumonia caused by HADV-5 infection, filling the current lack of effective treatment options in this field and possessing significant scientific research value and clinical application prospects.

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 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 discloses an epidermal skin model and a construction method and application thereof. The construction method comprises the following steps: performing gas-liquid culture on a transwell membrane on a 3D epidermal organ prepared from epidermal stem cells to prepare an epidermal skin model. The epidermis skin model prepared by the construction method can be rapidly amplified to realize large-scale modeling application in a short time, and the epidermis skin model is complete in differentiation, high in integrity, good in barrier function and beneficial to epidermis-related safety detection or efficacy detection of drugs or cosmetics.

A method for domesticating CHO-K1 cells into high-expression cells, comprising the following steps: culturing adherent CHO-K1 cells in a complete culture medium, to obtain log growth CHO-K1 cells; transferring the log growth CHO-K1 cells to a serum-free medium for suspension culture; performing a first liquid replacement domestication, to obtain domesticated CHO-K1 cells; placing the domesticated CHO-K1 cells in a chemically defined medium for suspension culture; performing a second liquid replacement domestication, to obtain domesticated suspended CHO-K1 cells; performing monoclonalization on the domesticated suspended CHO-K1 cells, to obtain CHO-K1 monoclonal cells; and screening the CHO-K1 monoclonal cells, to obtain high-expression CHO-K1 monoclonal cells. This method can domesticate CHO-K1 original parent cells into suspended cells, and screen cells having higher expression capability from the domesticated cell population, thereby improving expression levels.

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 utility model relates to the technical field of cell culture, and discloses a chamber structure adaptive to Faraday wave assembly, which comprises a pore plate, N mounting grooves are formed in the top surface of the pore plate, and N is greater than or equal to 1; the N container bodies are arranged on the pore plate in a rectangular array mode or a central symmetry mode and correspondingly arranged in the N mounting grooves, a plurality of culture grooves used for cell assembly are sequentially formed in the container bodies in the longitudinal direction from the top face to the bottom face, and the widths of the culture grooves are gradually decreased. According to the device, internal cells or cell microspheres can be gathered into a sound potential well of a standing wave sound field under the action of acoustic pressure, gravity and buoyancy under the action of the sound field and are tightly arranged according to an expected pattern, and a culture solution is longitudinally stacked in a plurality of culture tanks of which the widths are gradually reduced; longitudinal shape limitation on the tissue structure is realized, and complex three-dimensional longitudinal arrangement is constructed through mutual combination of the two. Therefore, the requirement of constructing any multi-scale and complex cell structure is met. And forming a multi-layer structure.

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 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.

To provide a cell preservation liquid and a cell preservation method which enable long-term refrigerated preservation of adherent cells while maintaining adherence capability to a culture substrate.SOLUTION: The present invention relates to: a preservation method for adherent cells which comprises refrigerated preservation of adherent cells in a non-frozen state in a cell preservation liquid that contains a high-molecular-weight; a cultivation method for adherent cells which comprises two-dimensional culturing of adherent cells after preservation via the preservation method; and a cell preservation liquid which includes a high-molecular-weight that is suitable for use in these methods.SELECTED DRAWING: None

The invention discloses a human NLRP7 mutation decidua type organ and application thereof, and belongs to the technical field of construction of type organs. The human NLRP7 mutant decidua organ is preserved in the China Center for Type Culture Collection, and is named as the human NLRP7 mutant decidua organ N7LLY (Homo sapiens), the preservation number is CCTCC (China Center for Type Culture Collection) NO: C2024375, and the preservation date is November 06, 2024. The human NLRP7 mutant decidua-like organ is applied to construction of a model for simulating an in-vivo NLRP7 mutant embryo decidua-like organ, and is applied to screening of drugs with prevention or treatment effects on grape embryo or embryo implantation failure. The human NLRP7 mutant decidua-like organ provided by the invention has endometrial epithelial cells and interstitial cells at the same time, expresses receptivity-related markers, and can retain pathological mutation and disease phenotypes of patients in vitro.

The invention relates to the technical field of reproductive biology, in particular to a method for efficiently constructing pig testis organs in vitro. Comprising the following steps: S1, enzymolysis of testicular tissues of piglets; and S2, testicular cell resuspension counting: uniformly suspending the testicular cells obtained after enzymolysis in the step S1, sequentially collecting the cells into a 96-well plate with a U-shaped bottom, and adding a DMEM base culture medium containing 5% of FBS, 10% of KSR and 1% of P / S into each well to complete resuspension culture of the cells. S3, testicular organ in-vitro culture: separating testicular cell masses, transferring the separated cell masses into a 96-well plate with a U-shaped bottom, and adding the cell masses into a DMEM base culture medium containing 5% of FBS, 10% of KSR and 1% of P / S for culture; and the quantity of cells in the 96-well plate with the U-shaped bottom is 5 * 104 cells per well. After the culture system is optimized, the experimental process is simplified, the testicular organs can be directly cultured in the 96-well plate with the U-shaped bottom, the replacement frequency is reduced, the pollution risk is reduced, the number of adopted cells is remarkably reduced, and the culture efficiency is improved.

The invention provides a construction method of a radioactive esophageal injury organ-like model. The construction method comprises the following steps: performing X-ray ionizing radiation on esophageal organs; wherein the esophageal organ comprises a rat esophageal organ, a mouse esophageal organ or a human esophageal organ; the radiation dosage is 0.5 to 4 Gy, and the radiation dosage rate is 1.5 to 2 Gy / min. Experimental results show that along with increase of irradiation dose, connection around spheres of human esophageal organs, mouse esophageal organs and rat esophageal organs is loose, cell debris is increased, and the sizes are smaller than those of normal organs at the same time. The construction of the radioactive esophageal injury organ-like model is of great significance for deeply understanding the injury mechanism of ionizing radiation to the esophagus, searching for effective protection and treatment measures and promoting the development of personalized radiotherapy.

The invention relates to the technical field of micro-fluidic intestinal organ chips, and particularly discloses a micro-fluidic intestinal organ chip as well as a construction method and application thereof. The chip body comprises an upper-layer chip, a porous film and a lower-layer chip; a porous film is arranged between the vein unit and the artery unit; the construction method comprises the following steps: (1) adding collagen into the intestinal culture unit, and inducing gel formation; (2) inoculating endothelial cells to the lower layer of the porous film; and (3) inoculating intestinal epithelial cells. The method can be applied to the simulation of the influence of the intestinal environment on the organism; the absorption of intestinal tracts on nutrient substances and toxins is simulated; the influence of the brain on the intestinal tract is simulated, and the influence of environmental changes (such as chronic pressure and depression) in the body on the intestinal tract is simulated through hormones, neurotransmitters, inflammatory factors or other bioactive substances in blood circulation on the structure and the barrier function of the intestinal tract; in-vivo simulation of immune and inflammatory reactions absorbed after digestion; and the kit can be used for replacing experimental animals in the drug screening process.

The invention belongs to the technical field of cell culture, and particularly relates to a method for promoting proliferation and differentiation of cervical squamous epithelial cells. The method comprises the following steps: S1, extracting and culturing primary cervix uteri squamous epithelial cells from cervix uteri tissues to obtain primary cervix uteri squamous epithelial cells; s2, adding the primary cervical squamous epithelial cells into a cell proliferation induction culture solution to induce cell proliferation, so as to obtain proliferated cervical squamous epithelial cells; and S3, uniformly mixing the proliferated cervical squamous epithelial cells with differentiation matrigel, and inducing cell differentiation by using a cell differentiation induction culture solution to obtain differentiated cervical squamous epithelial cells. Proliferation and differentiation of the cervical squamous epithelial cells are promoted by preparing a proper culture medium, and influence factors of proliferation and differentiation of the cervical squamous epithelial cells are researched by culturing the cervical squamous epithelial cells in vitro, so that the pathogenesis of cervical erosion and cervical cancer can be clarified; therefore, a direction is provided for research and development of medicines for treating cervical erosion and cervical cancer.

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 relates to a culture medium for high-density serum-free suspension culture of BHK-21 cells and application of the culture medium. The culture medium contains inorganic salt, amino acid, vitamins, energy substances, a buffer substance, an antioxidant, sterol, polyamine, an additive and an indicator, and the antioxidant comprises dihydrolipoic acid and S-acetyl-L-glutathione. According to the invention, the culture medium for serum-free suspension culture of the BHK-21 cells, which is clear in components and remarkable in culture effect, can be provided. The added dihydrolipoic acid and S-acetyl-L-glutathione are combined as antioxidants of the culture medium, so that the survival rate of BHK-21 cells cultured in the culture medium is improved while the oxidation resistance of the serum-free suspension culture medium is improved.

The present invention relates to novel lipid nanoparticles capable of liver-specific delivery of mRNA, a manufacturing method therefor, and a liver disease therapeutic agent capable of regenerating the liver by liver-specific delivery of the bio-reprogramming factor mRNA using same. The lipid nanoparticles according to the present invention allow for liver-specific delivery of mRNA at very high efficiency, and the lipid nanoparticles that encapsulate the mRNA-based bio-reprogramming factor mRNA are specifically delivered to liver tissues of an individual requiring liver regeneration to induce short-term expression, thereby easily setting the dose and expression period of a therapeutic agent, leading to a significant reduction in side effects of a therapeutic agent for liver regeneration using an existing cell reprogramming factor.

To provide a shellac modified object which can be changed from a state where cell adhesiveness to a mammalian cell is high, to a state where the cell adhesiveness is low.SOLUTION: A shellac modified object is expressed by a formula (1). In the formula, Z is a part excluding a carboxy group modified by a functional group Y in the structure of the shellac, X1 to X4 are a dialkylamino group, alkylamino group, hydroxy group, halogen group, H or the like.SELECTED DRAWING: Figure 1

The invention relates to a method for extracting high-purity extracellular vesicles from liver tissues of pelteobagrus vachelli, which comprises the following steps of: firstly, mixing collagenase II and collagenase IV, performing enzymolysis on the tissues, adding a CaCl2 solution, incubating, and digesting the liver tissues into a tissue suspension; then removing cells and fragments through low-speed centrifugation, and filtering to remove impurities by using needle filters of 0.8 [mu] m and 0.22 [mu] m; and finally, further purifying by combining ultracentrifugation with a high-precision iodixanol density gradient centrifugation method, and merging density layers with high purity to obtain high-purity EVs. The method has the characteristics of high extraction purity, high efficiency and simplicity and convenience in operation, and has a wide application prospect.

The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.

The present invention addresses the problem of providing: a method for preparing hepatic lineage cells in a large amount and with high purity; and a method for preparing liver system cells which can achieve high graft survival rate in a living body.　The present invention provides: a hepatoblast proliferation promoter and a culture medium, each of which contains a ROCK inhibitor, a TGFβ signaling pathway inhibitor, a Wnt signaling pathway activator, and a growth factor; and a proliferation / culture method using the hepatoblast proliferation promoter or the culture medium.

The present invention relates to glycoengineered extracellular vesicles, as well as methods of production of such glycoengineered extracellular vesicles, methods of glycoengineering extracellular vesicles, and uses of glycoengineered extracellular vesicles.

The invention belongs to the technical field of cell biology, and particularly discloses a culture medium and a culture method of fish primary liver cells. The culture medium of the fish primary liver cells comprises the following components: 90% of an M199 culture medium, 10% of fetal calf serum, 100 U / mL of penicillin, 0.1 mg / mL of streptomycin, 5 ng / mL of EGF, 10 [mu] U / ml of insulin and 10 mM of dexamethasone. Based on the culture medium, an efficient and stable fish liver cell culture method is developed by combining a standardized anesthesia and disinfection process, a precise dissection and liver sampling method, an optimized tissue digestion and cell separation scheme, an efficient cell purification and adherent culture system, a stable passage technology and the like; a standardized hepatocyte line establishment operation process is defined, and the problems that species binding exists in an existing fish liver cell culture method, and a standardized culture process suitable for multiple fish fingerlings is difficult to form are solved.

The invention discloses a cat kidney suspension culture cell line suitable for serum-free suspension culture and application of the cat kidney suspension culture cell line. The name of the cell line is F81-B4, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46350. After 50 generations of continuous subculture, the cells still can maintain the original proliferation level and cellular morphology, the cell number can reach 9.0 * 10 < 6 > / mL after the cells are subcultured at the density of 1.0 * 10 < 6 > / mL for three days, the multiplication time is 22-26 hours, and the cell proliferation speed is equivalent to that in a shake flask when the cells are amplified to a 10L bioreactor for culture. The cell line is highly susceptible to feline parvovirus, feline calicivirus, feline herpes virus, canine parvovirus and canine distemper virus, the virus content of a virus solution harvested 72 h after FPV virus inoculation can reach 107.5 TCID50 / mL, the virus content of a virus solution harvested 24 h after FCV virus inoculation can reach 1010.8 TCID50 / mL, the virus content of a virus solution harvested 48 h after FHV-1 virus inoculation can reach 107.5 TCID50 / mL, compared with an existing F81 adherent cell production process, the production process has the advantages that the production efficiency is high, and the production cost is low. The method has the characteristics of rapid lesion, short virus collection time and high titer. The cell matrix is an ideal cell matrix which can be used for culturing the three viruses.