138 results about "Cellular engineering" patented technology

Disclosed are systems and methods for improving processes for developing cell therapies by applying machine learning to data including manufacturing process data and clinical measurements (e.g., patient response and treatment data) to determine parameters and settings for a manufacturing process for engineering cells for use in cell therapy. Parameters and settings for a manufacturing process for genetically engineered T-cells including, but not limited to, Chimeric Antigen Receptor (CAR) T cells can be determined. A method can include receiving a set of process parameters of a cell engineering process, predicting a clinical response associated with an output of the cell engineering process by applying a machine learning model on the received set of process parameters, where the machine learning model is trained on process parameter data and clinical response data, and generating a visualization for use in a graphical user interface of the predicted clinical response.

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 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.

Cell engineering, soft robotics and wearable electronics often desire soft materials that are easy to deform, self-heal, and relax stress. Hydrogel, a type of hydrophilic networks, which can be made responsive to environmental stimuli, are often used in the aforementioned applications. However, conventional hydrogels often suffer from poor stretchability and repairability. Here, we report hydrogels consisting of boronic ester dynamic covalent bonds in a double network of poly(vinyl alcohol) together with chitosan, demonstrating extreme stretchability (up to 310 times of the original length), instant self-healing (within 5 sec), reusability as well as inherent adhesion. Their instant stress relaxation as a result of low activation energy of the boronic ester bond exchange (less than 20 KJ / mol) contributes to the extreme stretchability and self-healing behaviors. The hydrophilic environment readily incorporates various additives and provide opportunities in conductive soft materials, bio-signaling, and soft artificial joint.

The present application relates to the field of plant tissue culture technology, and particularly discloses a lily stem cell abundance ex vivo regulation method. The lily bulb is used as the starting material, stem cell magnitude amplification and regulation culture can produce lily tissue organ materials with high stem cell abundance. The present application provides sterile lily materials with high stem cell abundance, which has the characteristics of strong regenerative ability, high proliferation rate, etc., is the basis for protoplast preparation and high-efficiency regeneration, and is suitable for different lily varieties, and provides technical support for cell engineering breeding, seedling scale breeding, etc.

The invention relates to the field of cell engineering and gene engineering, in particular to a method for enhancing the multiplication capacity of umbilical cord mesenchymal stem cells and application. The expression or activity of the STN1 gene in the umbilical cord mesenchymal stem cells is reduced through an accelerant, and the umbilical cord mesenchymal stem cells with enhanced multiplication capacity are obtained. By knocking down the hUC-MSCs of STN1, the proliferation speed of the hUC-MSCs is higher, DNA replication is more active, the clone formation ability is higher, and the aging proportion of the hUC-MSCs can be remarkably reduced.

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

The invention belongs to the field of cell biology, and particularly relates to a preparation method and application of an extracellular matrix of cell-engineered human collagen. The preparation method comprises the following steps: S1, culturing human induced pluripotent stem cells or a cell culture comprising the human induced pluripotent stem cells, and directly inducing differentiation to obtain induced mesenchymal stem cells or a cell culture comprising the induced mesenchymal stem cells; and S2, carrying out 3D culture on the induced mesenchymal stem cells through a 3D porous scaffold carrier. The extracellular matrix prepared by the preparation method provided by the invention is rich in collagen, so that the mesenchymal stem cells can be better promoted to be differentiated into osteoblasts, and meanwhile, a new thought is provided for the preparation of the collagen.

In one aspect, a method of treating an NK cell to induce a gene edit. The method may include contacting an NK cell with a transfection composition including poly-L-glutamic acid (y- PGA), a single-stranded oligo deoxynucleotide (ssODN) enhancer, or a combination thereof; contacting the NK cell with an editing composition including: a DNA-PK inhibitor, and an HD AC inhibitor; electroporating the NK cell; and contacting the NK cell with a recovery composition including DNase I. In another aspect, a method of preparing an engineered NK cell comprising a fluorescent reporter at an endogenous locus. In another aspect, a method of treating an NK cell to introduce an exogenous nucleic acid.

The present disclosure relates to immunotherapy. In more specific embodiments, the present disclosure provides systems, compositions, methods and uses of viral vectors comprising nucleic acid sequence of interest that encodes at least one therapeutic product, and a nucleic acid sequence encoding at least one nuclease, for in vivo targeted insertion of the nucleic acid sequence of interest into a target locus within at least one cell of the T lineage.

Disclosed are systems and methods for improving processes for developing cell therapies by applying machine learning to data including manufacturing process data and clinical measurements (e.g., patient response and treatment data) to determine parameters and settings for a manufacturing process for engineering cells for use in cell therapy. Parameters and settings for a manufacturing process for genetically engineered T-cells including, but not limited to, Chimeric Antigen Receptor (CAR) T cells can be determined. A method can include receiving a set of process parameters of a cell engineering process, predicting a clinical response associated with an output of the cell engineering process by applying a machine learning model on the received set of process parameters, where the machine learning model is trained on process parameter data and clinical response data, and generating a visualization for use in a graphical user interface of the predicted clinical response.

The invention belongs to the field of biosynthesis of macrocycloalkanone compounds, and particularly relates to a biosynthesis method and application of macrocycloalkanone compounds based on a cell engineering technology. The method comprises the following steps: culturing gland cells in a culture medium, adding reaction substrates aldehydes and fatty acid, and synthesizing the macrocycloalkanone compound from the aldehydes and fatty acid through cyclization reaction in the cells. The method provided by the invention overcomes the defects of complex process, low yield and high production control difficulty in artificial synthesis, and extremely low yield and unstable product properties in natural product extraction, and has the beneficial effects of realizing large-scale and engineering production, reducing the production cost and improving the yield and quality.

The invention provides a composition, cell culture and application of the composition in differentiation of human pluripotent stem cells to myocardial cells and / or construction of human-derived heart organoid, and belongs to the technical field of cell engineering. The invention provides a composition. The composition comprises a fatty acid receptor antagonist, a fatty acid synthetase inhibitor and a fatty acid oxidation activator. The composition provided by the invention can regulate cell metabolism, breaks through the contradiction that the proliferation capacity and the cell maturity cannot be obtained at the same time in the traditional technology, can maintain the myocardial cells in a proliferation stage, and obtains the myocardial cells with relatively mature structures and functions, and the contractility of 3D heart organs constructed by the myocardial cells is greatly improved. The invention provides a high-quality cell source for myocardial regeneration treatment, high-throughput drug cardiotoxicity screening and heart disease mechanism research, and has remarkable clinical application value and industrialization prospect.

This invention discloses an immune cell that secretes type 2 cytokines under hypoxic conditions and its applications, belonging to the fields of genetic engineering and cell engineering. The invention first constructs a nucleotide fragment encoding hypoxia-induced type 2 cytokines, including a signal peptide gene sequence, a CAR molecule nucleotide sequence targeting CD19 and CD22, and an IL-4 or IL-10 nucleotide sequence containing multiple HRE motifs of a hypoxia-inducible promoter. This nucleotide fragment is delivered to immune cells for stable expression, yielding an immune cell that secretes type 2 cytokines under hypoxic conditions. This immune cell secretes IL-4 or IL-10 only under hypoxic conditions, thereby leveraging the metabolic reprogramming effect of IL-4 or IL-10 on immune cells to enhance their activity under hypoxic conditions, thus achieving better long-term anti-tumor effects.

The application belongs to the technical field of animal cell engineering, and particularly relates to a pigeon fibroblast cell strain X4 and application thereof. A pigeon fibroblast cell strain is provided, which is named pigeon fibroblast cell X4, and the preservation number is CGMCC No. 46754. The pigeon fibroblast cell strain can be used for virus isolation or amplification, virus pathogenic mechanism research, and development of antiviral drugs and vaccines, and has a good application prospect in pigeon disease research.

The invention provides a larimichthys crocea head kidney macrophage line as well as a construction method and application thereof, belongs to the technical field of cell engineering, and particularly provides the larimichthys crocea head kidney macrophage line which is named as LYC-hK and has the preservation number of CCTCC NO: C2025234. The larimichthys crocea head kidney macrophage line is constructed through the steps S1-S6, and the construction method of the larimichthys crocea head kidney macrophage line is applied to the field of cell engineering. The larimichthys crocea head kidney macrophage line provided by the invention can be used for research on pathogenic infection and pathogenic mechanism of larimichthys crocea, research on anti-infection immune response of larimichthys crocea and research on exogenous gene expression.

Provided herein, inter alia, are methods and compositions for engineering T cells. The methods include culturing a T cell with insulin during engineering of the T cell. The methods provided herein are contemplated to increase cell viability, expansion and gene editing efficiency, thereby allowing an increase in the total number of engineered T cells.

The invention belongs to the cross technical field of cell engineering, nano-drugs and biological manufacturing, and particularly discloses a drug-loaded vesicle based on denucleated cells as well as a preparation method and application of the drug-loaded vesicle. According to the invention, a cell suspension and a cytoskeleton relaxant are co-incubated to relax an actin skeleton and weaken nucleoplasm connection; then loading the treated cell suspension on a multi-layer discontinuous density gradient centrifugal medium, realizing physical separation of cell nucleuses and cytoplasm through high-speed centrifugation according to buoyancy density difference, and collecting components of a specific interface to obtain high-purity denucleated cells with a complete membrane structure; then co-incubating the denucleated cells and ROS response type lipidosome (co-carrying therapeutic siRNA and a sound-sensitive agent Ce6) prepared in advance, so that the lipidosome is wrapped or anchored by a denucleated cell membrane; finally, the composite system is subjected to extrusion treatment through a microporous membrane, the drug-loaded vesicles uniform in particle size and stable in structure are obtained, and the drug-loaded vesicles are suitable for various application scenes such as anti-tumor treatment and RNA vaccine delivery.

The application belongs to the technical field of biology, and relates to an anti-H10 subtype influenza virus hemagglutinin protein monoclonal antibody and application thereof. The application is obtained by using cell engineering and antibody engineering technology, and is a hybridoma cell line secreting the anti-hemagglutinin protein monoclonal antibody. The anti-hemagglutinin protein monoclonal antibody 1E1 is prepared by inducing ascites of the same strain mouse, and is identified as IgG2a and kappa type. The application of the antibody is realized by affinity purification and immunization methods. The application can specifically recognize the hemagglutinin protein of the H10 subtype influenza virus. The heavy chain variable region amino acid sequence of the antibody is shown in SEQ ID No. 2, and the light chain variable region amino acid sequence is shown in SEQ ID No. 4. The application provides an effective tool for auxiliary diagnosis of H10 subtype influenza virus infection in clinical samples, and can be applied to various detection technologies and clinical and experimental researches.

The application provides a method for constructing an immortalized bovine nasopharyngeal tonsil epithelial cell line and application, and belongs to the technical field of cell engineering. The healthy yellow bovine nasopharyngeal tonsil is used as a material, and the primary epithelial cells are separated by enzyme digestion. The 3rd to 4th generation cells are taken, infected with SV40 T lentivirus (MOI=80), and screened by 1.5 μg / mL puromycin to obtain a stable immortalized cell line. The cell line is continuously passaged to the 50th generation and still stably expresses SV40LT protein and epithelial specific markers CD326, CD324 and KRT8, maintains a typical pavement stone-like morphology, normal proliferation and no malignant transformation. Infection experiments show that the cell is highly sensitive to FMDV, supports efficient replication, and the virus titer can reach 10 7.5 TCID 50 / mL. The application overcomes the defects of the existing immortalized bovine nasopharyngeal tonsil epithelial cell model, such as limited source, large batch difference, low sensitivity and poor persistence, and provides a stable and reliable in vitro model for the research of foot-and-mouth disease prevention and control related mechanisms, drug screening and vaccine development.

The invention belongs to the technical field of animal reproductive biology and cell engineering, and particularly relates to a fluorescence labeling expression system for visualizing an oocyte nucleus maturation process and application of the fluorescence labeling expression system. The fluorescence labeling expression system comprises an eukaryotic expression vector pVenus-LMNB1; the eukaryotic expression vector pVenus-LMNB1 comprises a nucleotide sequence for coding the fusion protein Venus-LMNB1. According to the invention, a nuclear fiber layer protein stably expressed on a nuclear membrane is selected as a research object, a eukaryotic expression vector pVenus-LMNB1 is constructed by using a molecular cloning technology, an LMNB1 gene containing a green fluorescent label Venus is overexpressed in a porcine oocyte by using a microinjection technology, fluorescent protein labeling of a structural protein on the nuclear membrane is utilized, and the expression vector pVenus-LMNB1 is constructed. The expression positioning condition of the LMNB1 in the maturation process of the porcine oocytes is observed, and a powerful tool is provided for development of in-vitro culture related researches of the porcine oocytes.

The present invention relates in general to a platform to selectively eliminate unwanted cells from a population of cells. The invention relates to self-amplifying RNA constructs for cell purification preferably during differentiation and cell engineering.

To provide an automated method of producing viral vectors, utilizing engineered viral vector-producing cell lines, or packaging cells, within a fully-enclosed cell engineering system.SOLUTION: A method for automated production of a viral vector, includes: introducing an engineered viral producer cell into a high-temperature chamber of a fully enclosed cell engineering system; transducing the engineered viral producer cell with a vector encoding a gene of interest to produce a transduced viral producer cell; expanding the transduced viral producer cell and producing the viral vector within the cell; transferring the expanded producer cell to a downstream processing module; and isolating the viral vector and purifying the viral vector, the above-described steps being performed in a closed and automated process. Exemplary viral vectors that can be produced include lentivirus vectors, adeno-associated virus vectors, baculovirus vectors and retrovirus vectors.SELECTED DRAWING: Figure 1

The present invention provides a cell suitable for gene engineering, cell engineering and cellular medicine, and a method for producing the same. The present invention provides a technique for removing repetitive sequences present in a specific gene region in two or more alleles, thereby facilitating gene targeting or sequencing in the region.

The invention belongs to the technical field of cell engineering and genetic engineering, and particularly relates to application of an FBXO32 gene in regulation and control of skeletal muscle satellite cell proliferation and feed utilization rate. According to overexpression of the FBXO32 gene in skeletal muscle satellite cells, it is found that the mRNA expression quantity of skeletal muscle satellite cell proliferation marker genes KI67 and CCND1 is remarkably increased, and the protein expression level is also remarkably improved. CCK-8 and EdU detection results also show that the FBXO32 gene or the encoded protein thereof also has a promotion effect on skeletal muscle satellite cell proliferation. The overexpressed FBXO32 gene or the encoded protein thereof can promote the proliferation of skeletal muscle satellite cells, and lays a foundation for improving meat traits of livestock and poultry, improving the feed utilization rate of the livestock and poultry and improving the meat traits of the livestock and poultry.

The patent discloses a method for quickly establishing a Cabernet Sauvignon grape pericarp callus regeneration system, and belongs to the technical field of cell engineering. The method takes the pericarp of Cabernet Sauvignon grape as an explant, realizes efficient induction and rapid regeneration of callus through three key steps of explant disinfection treatment, primary callus induction culture, callus proliferation and subculture. The method optimizes the culture medium formula, adds specific concentrations of 6-BA, KT, IAA, NAA and 2,4-D in the induction medium, and significantly improves the induction rate and growth rate of callus. Experimental results show that the stable callus regeneration system can be obtained in 40-50 days by using the method, the callus induction rate can reach more than 90%, and the survival rate of regenerated plants is high. The patent has the advantages of simple operation, good repeatability and short cycle, and provides an effective technical means for genetic improvement, gene function research and rapid propagation of wine grapes, and has important theoretical value and application prospect.

The invention provides a mesenchymal stem cell cryopreservation solution and a preparation method thereof, and belongs to the technical field of cell engineering. According to the cryopreservation liquid, diluted DMEM is used as a basic culture medium, and trehalose, L-proline, polyethylene glycol, sodium carboxymethyl cellulose, sodium glycinate, sericin, glycerin, resveratrol and other components are introduced as additives on the basis of the basic culture medium. The formula of the cryopreservation liquid adopts a DMSO-free and serum-free scheme, sodium glycinate, sericin and glycerol are used as composite protective agents, ice crystal damage can be effectively relieved, osmotic pressure balance can be maintained, cell membrane and protein structures can be protected, the effect is better than that of DMSO and glycerol, and cytotoxicity is avoided. In addition, resveratrol, coryne, astragaloside and other components are used as growth factors, so that the activity of the mesenchymal stem cells in a cryopreservation state is maintained. The survival rate of the umbilical cord mesenchymal stem cells cryopreserved for 2 years can reach 99.83% at most, the cell viability OD value reaches 0.96, and the cell adherence and growth conditions are good when the cells are observed under a microscope.

The invention provides a drug combination composition for in-vivo target cell engineering modification, and target cells do not include myeloid cells. The drug combination composition comprises a first drug and a second drug, wherein the active ingredient of the first drug is a substance capable of saturating, inhibiting or eliminating phagocytic ability of in-vivo myeloid cells, and the active ingredient of the second drug is an in-vivo target cell capable of being engineered. Through combined application of the myeloid cell inhibition drug and the cell engineering modification drug targeting the target cells, endocytosis of the myeloid cells to the target drug is reduced or eliminated before engineering modification of the target cells, so that the off-target effect is reduced, the lymphatic organ targeting property can be improved, and the treatment effect of the myeloid cells is improved. The toxic and side effects of cell engineering modification drugs on the liver are reduced, and the method has important significance on in-vivo (in-situ) lymphocyte engineering modification (including gene editing) or immunotherapy (such as in-vivo CAR T cell therapy).

The application belongs to the technical field of cell engineering, and particularly relates to a bispecific antibody targeting CD22 and CD19, a chimeric antigen receptor thereof and application. In the bispecific antibody, a CD22 light chain and a CD22 heavy chain are connected through a linker 4 to form a CD22 single-chain antibody, and a CD19 light chain and a CD19 heavy chain are connected through a linker 4 to form a CD19 single-chain antibody, which comprises one of the following structures: (1) the CD22 single-chain antibody and the CD19 single-chain antibody are connected through a linker 1 or a liner 2; (2) the two ends of the CD19 single-chain antibody are connected with the CD22 light chain and the CD22 heavy chain through a linker 3, respectively. The chimeric antigen receptor provided by the application can simultaneously target two different tumor antigens, improve the killing of tumor cells, reduce the probability of immune escape, and reduce the tumor recurrence rate after CAR-T treatment.

This invention belongs to the field of cell engineering technology, specifically relating to an engineered mitochondrial nanoshield, its preparation method, and its applications. The engineered mitochondrial nanoshield can resist damage from high concentrations of calcium ions in the extracellular environment, and also possesses BMMSCs targeting ability and ROS-responsive characteristics. After intravenous injection, the engineered mitochondrial nanoshield can selectively target BMMSCs to alleviate cellular senescence and increase bone mass in aged mice.