2662results about "Vertebrate cells" patented technology

The disclosure provides compositions and methods for the preparation, manufacture and use of an adeno-associated virus (AAV) particle for the vectorized delivery of an antibody molecule that binds to tau.

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.

Allografts for soft tissue repair, including breast reconstruction and other plastic surgery procedures, are disclosed. In an exemplary embodiment, first and second cuts are made into the reticular layer of a skin sample to separate an improved acellular dermal matrix (ACDM) which forms a dermal tissue form useful as a graft. The dermal tissue form consists essentially of reticular dermis and has a first exposed surface formed by reticular dermis, an opposite second exposed surface formed by reticular dermis, and has substantially uniform density and uniform porosity between the first and second exposed surfaces, which allows and promotes rapid and efficient cellular ingrowth and tissue ingrowth substantially equally from either the first or second exposed surfaces upon implantation.

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 relates to a screening method for odor-suppressing materials. A screening method according to the present invention includes: causing a test substance and an odor-causing substance to act on at least one olfactory receptor polypeptide selected from the group consisting of olfactory receptor OR2AG2 and polypeptides that include an amino acid sequence having 80% or greater identity with the amino acid sequence of the aforementioned olfactory receptor and exhibit responsiveness to odor-causing substances; measuring the response of the olfactory receptor polypeptide to the odor-causing substance; and, on the basis of the measured response, identifying test substances that suppress a response of the olfactory receptor polypeptide as candidate substances for an odor-suppressing material.

The invention discloses application of an MDH2 inhibitor in preparation of a preparation for reducing toxicity of fumonisins, and belongs to the technical field of biology. A fumonisins treatment experiment is carried out on MDH2 gene mutant cells and wild type MDH2 gene cells, and the result shows that the cell activity of the MDH2 gene mutant cells is remarkably improved by 15.2% compared with that of wild type MDH2 gene expression cells under the condition that the fumonisins concentration is 100 micrograms / mL, the cell apoptosis rate is remarkably reduced by 7.76%, and the cell apoptosis rate can be reduced by 7.76%. Therefore, the MDH2 gene mutation can improve the tolerance to the fumonisins. In addition, the LW6 inhibitor is used for inhibiting the MDH2 activity of the wild type IPEC-J2 cell, and the tolerance of the cell to the fumonisins can also be improved. The invention provides a potential solution way for preventing adverse reaction caused by fumonisins, and provides a theoretical basis for preventing and controlling harm of FB1 to food and animal husbandry.

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.

Provided herein are transposon / transposase systems for transposing and selecting of transposed cells where the system includes a transposase or polynucleotide encoding a transposase; and an exogenous nucleic acid sequence comprising: a transposon comprising: a 5' inverted terminal repeat (ITR), a first polynucleotide encoding a gene of interest; and a 3' inverted terminal repeat (ITR); and (ii) a polynucleotide encoding an at least first selection marker located outside of the 5' ITR and 3' ITR, where upon introducing (a) and (b) into a cell the transposon is transposed into the cell's genome and the polynucleotide encoding the selection marker is episomal.

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 application of a milk exosome in preparation of a medicine for resisting porcine epidemic diarrhea virus, and belongs to the technical field of biological medicine, and the milk exosome is prepared by adopting a citric acid dissolved protein method and an ultrafiltration concentration method. The invention further discloses a medicine for resisting the porcine epidemic diarrhea virus, and the medicine contains the milk exosome in the application. According to the present invention, the porcine viral diarrhea virus resistance of the milk exosome milk-exo is researched, the results show that the milk-exo can significantly inhibit the replication ability of the PEDV virus, and can reduce the damage of the PEDV virus to PK1 cells and Vero cells, such that the application effect of the milk-exo in the preparation of the porcine epidemic diarrhea virus resistance drug is proved. The preparation method of the exosome from milk is simple, the raw materials are easy to obtain, and the exosome has a good application prospect.

The invention discloses an effect of an APOA gene in relieving vomitoxin injury of bovine liver cells by vitamin E, and relates to the technical field of the effect of vitamin E in relieving vomitoxin injury of bovine liver cells. The APOA gene refers to an APOA1 gene, an APOA2 gene and an APOA4 gene; the CDS sequence of the APOA1 gene is as shown in SEQ ID NO. 19; the CDS sequence of the APOA2 gene is as shown in SEQ ID NO. 20; the CDS sequence of the APOA4 gene is as shown in SEQ ID NO. 21; the effect is that the vitamin E regulates an APOA gene to be enriched to a cholesterol metabolic pathway. According to the effect of the APOA gene in relieving vomitoxin-caused cattle liver cells, it is found that the vitamin E can effectively protect the cell membrane integrity of the cattle liver cells and can relieve cell viability decline, oxidative stress, inflammatory response and cell apoptosis caused by vomitoxin; vitamin E regulates the cholesterol metabolic pathway by regulating the APOA gene level so as to play a role in protecting liver cells of dairy cows.

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 separating an exosome from a heterologous expression cell or a primary culture cell, and a use of an exosome in biological analysis, relating to the technical field of biomedicine. The method for separating the exosome from the heterologous expression cell or the primary culture cell of the present invention comprises: carrying out differential centrifugation on a solution to be extracted that contains an exosome, then taking out a first supernatant, carrying out high-speed centrifugation again, respectively adding iodixanol into an obtained second supernatant and a precipitate, and carrying out spontaneous density gradient centrifugation to obtain a separated purified exosome. According to the present invention, an iodixanol spontaneous density gradient centrifugation method is innovatively used to successfully carry out separation and purification to obtain the exosome having high purity and high yield, and the separated exosome is used as an experimental development universal platform for researching a membrane protein structure and functional characteristics in an original ecological cell membrane environment, and is used for biological analysis, drug screening, and disease diagnosis and treatment.

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 human-derived iPSC (induced pluripotent stem cell)-induced liver organ and a preparation method thereof. The method comprises the following steps: digesting human induced pluripotent stem cells obtained by culture into single cells; single cells are prepared into cell clusters through self-balling, or cell matrigel microspheres are prepared by using a T-type microfluidic technology and then are distributed one by one, so that one cell cluster is formed in each pore of the low-adsorption 96-pore plate; carrying out step-by-step induction culture on the obtained cell cluster, and culturing in a culture medium I for 5 hours; replacing with a culture medium II and culturing for 2 days; a culture medium III is used for culturing for 2 days; replacing with a culture medium IV and culturing for 5 days; replacing with a culture medium V, and culturing for 6 days; and replacing with a culture medium VI, and culturing for 10 days to obtain the mature human liver organ. According to the invention, liver organs with human normal liver cell characteristics and protein expression can be formed within 25 days, and the method can realize homogenization and high efficiency of iPSC induction.

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.

To provide a cell injection device, a cell collection accessory jig, and a cell collection method capable of stably injecting foreign cells into a cell mass regardless of the skill and technique of an operator.SOLUTION: A cell injection device includes: a cell collection accessory jig 24 including a filter section 26 having bubbles of a size that allows a liquid medium D to pass through but not cells Ce; an operation manipulator equipped with an injection pipette 90 capable of collecting and discharging a plurality of cells Ce and the liquid medium D from an opening 90a at the tip; an injection pump capable of adjusting the internal pressure of the injection pipette 90; and a controller that, after collecting a plurality of cells Ce and the liquid medium D inside the injection pipette 90, applies positive pressure to the inside of the injection pipette 90 while pressing the opening 90a at the tip of the injection pipette 90 against the filter section 26, thereby discharging the liquid medium D through the filter section 26 and aligning the exogenous cells Ce inside the injection pipette 90 in rosary-like pattern.SELECTED DRAWING: Figure 2

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.

Disclosed herein are compositions of transcription activator-like effectors transcription factors and methods of using the compositions for inducing gene expression of mammalian genes.

The invention relates to the technical field of medicines and biology, in particular to a mouse EGFR (Epidermal Growth Factor Receptor) mutant lung adenocarcinoma cell line (EP Mus musculus) as well as preparation and application thereof. The invention provides an EP cell line ([EGFRE19delta (mutation); trp53 (deletion), EP] is derived from a lung adenocarcinoma model of a C57BL / 6 strain mouse, carries the most common EGFRE19del mutation in lung adenocarcinoma, is a first lung adenocarcinoma mouse cell line carrying EGFR mutation known in the world at present, has the in-vivo tumorigenesis ability of the C57BL / 6 strain mouse, and fills the blank of the mouse EGFR mutant cell line. The cell line provided by the invention has the background of EGFRE19del homozygous activation mutation and Trp53 homozygous deletion mutation, is sensitive to EGFR-TKI targeted therapy, can be used for in-situ tumor formation in subcutaneous and lung of an immune normal mouse C57BL / 6, and is suitable for immunotherapy and targeted therapy research of EGFR mutation lung adenocarcinoma.

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

To provide hepatocyte organoids derived from stem cells such as iPS cells that retain the characteristics of hepatocytes and can be cultured for a long period of time, and to provide methods for producing hepatocyte cell lines containing the same.SOLUTION: The present invention provides a method for producing a hepatocyte cell line that includes a hepatocyte organoid, comprising a step of culturing cells derived from stem cells in a medium containing an intranuclear receptor agonist or a Wnt / β-catenin pathway activator. The present invention also provides a hepatocyte organoid derived from a stem cell, which can be cultured for 21 days or more.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.