264 results about "Cellular engineering" patented technology

Provided herein are compositions and methods for improving non-viral insertion of genes into safe harbor loci of cells such as immune cells.

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 discloses application of a Wnt2 gene in buffalo ovarian granular cells, and belongs to the technical field of cell engineering and gene engineering. Wnt2 is taken as a research object, the Wnt2 overexpression vector or Wnt2 siRNA is transfected to the ovarian granulosa cells respectively by constructing the Wnt2 overexpression vector and synthesizing small interfering RNA (Wnt2 siRNA), then changes of signal channel gene mRNA and protein level related to ovarian granulosa cell apoptosis, proliferation and E2 and P4 secretion are detected, and finally phenotypic changes of the ovarian granulosa cells are detected. Results show that Wnt2 can inhibit apoptosis of ovarian granular cells and promote proliferation and E2 and P4 secretion. By exploring the application of the Wnt2 in the ovarian granular cells, the application has good application in researching an ovarian follicle atresia mechanism, initial estrus initiation and the like.

The invention provides a method for enhancing Treg cell functions and application, and belongs to the technical field of cell engineering. According to the method, hUC-MSCs are irradiated by gamma rays to form feeder layer cells, and then the feeder layer cells and Treg cells are co-cultured to obtain the Treg cells with enhanced cell functions. Through the co-culture of the feeder layer cells and the Treg cells, the inhibition ability of the Treg cells on activation and proliferation of immune cells can be enhanced, and the feeder layer cells and the Treg cells can be used as universal cells in clinical researches of autoimmune diseases, graft versus host diseases and the like.

The invention discloses a serum-free culture medium for in-vitro proliferation of poultry muscle stem cells and application of the serum-free culture medium, and belongs to the technical field of cell engineering, a basic serum-free culture medium is DMEM-F12, a complete serum culture medium is a mixture of DMEM-F12 added with 10% of fetal calf serum and 1% of mycillin, and the volume ratio of a culture medium supplement to the basic culture medium is 1: 1000; the serum-free culture medium contains a supplement; the supplement comprises the following compositions: insulin, transferrin, fibroblast growth factor 2, epidermal growth factor, catalase, ethanolamine, vitamin B12, linoleic acid, linolenic acid, retinol acetate, tocopherol, L-alanine, L-proline, sodium selenite, sodium pyruvate, reduced glutathione, berberine hydrochloride and analogues thereof.

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 an SFRP1 gene in buffalo ovarian granular cells, and belongs to the technical field of cell engineering and gene engineering. According to the invention, SFRP1 is taken as a research object, an SFRP1 overexpression vector or siRNA-SFRP1 is respectively transfected to ovarian granular cells by constructing the SFRP1 overexpression vector and synthesizing small interfering RNA (siRNA-SFRP1), then changes of signal channel gene mRNA and protein level related to ovarian granular cell apoptosis, proliferation and E2 and P4 secretion are detected, and finally phenotypic changes of the ovarian granular cells are detected. Results show that the SFRP1 can inhibit apoptosis of ovarian granular cells and promote proliferation and E2 and P4 secretion. The application of the SFRP1 in the ovarian granular cells is explored, and the SFRP1 has good application in research of an ovarian follicle atresia mechanism, initial estrus initiation and the like.

The invention belongs to the technical field of cell engineering and biochemical detection, and particularly relates to an erythrocyte membrane with a complete structure and a preparation method and application thereof. The red blood cells are split by limiting the components of the release liquid, the conditions are mild, protein components such as hemoglobin in the red blood cells can be released, the cell attributes are removed, the integrity of the internal skeleton structure and the activity of the membrane surface antigen are reserved, the complete structure of the red cell membrane is kept, meanwhile, the key blood group antigen on the surface is reserved, and the effect is good. The problem of antigenicity reduction caused by erythrocyte membrane fragmentation is avoided. After the erythrocytes are lysed, the nanoparticles are coupled to the surface of the erythrocyte membrane, so that the agglutination reaction of the erythrocyte membrane under the induction of the antibody can be visualized. The engineering red blood cell biological membrane is preserved by adopting a fixed program, so that the damage of ice crystals to the engineering red blood cell biological membrane can be effectively avoided, the antigenicity of the membrane is not influenced, the membrane can be preserved for a long time at normal temperature, and the shelf life can be prolonged to two years or more.

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 is applicable to the technical field of cell engineering, and provides a mitochondria-enhanced dental pulp mesenchymal stem cell and a preparation method and application thereof, and the preparation method comprises the following steps: co-culturing platelet-rich fibrin and dental pulp mesenchymal stem cells by using a Transwell chamber to obtain the mitochondria-enhanced dental pulp mesenchymal stem cell. The platelet-rich fibrin transmits mitochondria to the dental pulp mesenchymal stem cells to improve the regeneration capacity of the dental pulp mesenchymal stem cells. According to the invention, an autologous platelet enrichment, namely platelet-rich fibrin, is used as a mitochondria donor with a respiratory function, mitochondria is continuously transferred to the dental pulp-derived mesenchymal stem cells through a cell bioengineering technology, and the mitochondria-enhanced dental pulp mesenchymal stem cells are constructed and applied to tissue damage repair; the vascular regeneration regulation and control capability of the dental pulp mesenchymal stem cells is obviously improved, and the existing mesenchymal stem cell treatment technology is obviously improved.

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 invention relates to the field of cell engineering, in particular to application of CDH1 in promoting cell production and / or exosome release. According to the invention, after the lentiviral vector of the over-expressed CDH1 gene is constructed, tumor cells are transfected by the lentiviral vector, and finally the tumor cell strain capable of stably expressing CDH1 is obtained. Through extraction and detection, the cell strain obviously improves the yield of the exosome in unit cell and unit culture medium volume, and the yield of the exosome is obviously improved.

Methods are provided for reprogramming M2-like macrophages to M1-like macrophages, which reverses the proinflammatory to anti-inflammatory shift observed during the course of certain cancers, co-administered with one or more types of engineered cells such as, without limitation, CAR T-cells, engineered natural killer cells, engineered stem cells or the like. The compounds comprise an immune modulator that targets a pattern recognition receptor of a cell and are specific to the cells of interest through the incorporation of a targeting moiety (e.g., folate or a functional fragment or analog thereof). Releasable and / or non-releasable linkers can be included and engineered to facilitate the optimal delivery of the immune modulator. The compounds and compositions can be employed in one or more methods of treatment for cancers.

The invention relates to the field of cell engineering, in particular to application of CD40 in promoting cell production and / or exosome release. According to the invention, after a lentiviral vector overexpressing the CD40 gene is constructed, the lentiviral vector transfects tumor cells, and finally the tumor cell strain stably expressing CD40 is obtained. Through extraction and detection, the cell strain obviously improves the yield of the exosome in unit cell and unit culture medium volume, and the yield of the exosome is obviously improved.

The invention relates to the technical field of biological medicine, and provides a ginseng cell engineering tumor vaccine as well as a preparation method and application thereof. The ginseng cell engineering tumor vaccine comprises an exosome derived from a recombinant ginseng cell capable of expressing a tumor antigen; the recombinant ginseng cell capable of expressing the tumor antigen is obtained by introducing a recombinant plasmid containing tumor antigen DNA into a ginseng cell. According to the invention, ginseng cells in Chinese herbal medicines are taken as host cells for recombinant expression of tumor antigens, large-scale production of tumor vaccines can be realized through construction of an engineering recombinant cell system, biocompatibility and uniformity of the vaccines can be maintained, and safety risks can be reduced. Meanwhile, the prepared tumor antigen recombinant expression ginseng cell exosome can integrate the antigen, an adjuvant and a carrier, and the antigen phagocytosis, maturation and antigen peptide presentation capabilities of APCs are enhanced.

The invention provides a preparation method and application of CAR-T cells for overexpressing C1QBP, and relates to the field of cell engineering. According to the CAR-T cell overexpressing the C1QBP, the energy metabolism of the cell is optimized by improving the oxidative phosphorylation of mitochondria, the formation of memory-like T cells is promoted, and the anti-tumor activity and long-term durability of the CAR-T cell are enhanced. The cells can survive in a tumor microenvironment for a long time, and the anti-tumor effect of the cells is more durable than that of traditional CAR-T cells. Therefore, a new direction is provided for research and development of cellular immunotherapy.

The invention relates to the technical field of cell engineering, in particular to a breast cancer carboplatin drug-resistant cell strain and a construction method thereof, the breast cancer carboplatin drug-resistant cell strain is preserved in Guangdong Microbial Culture Collection Center on October 16, 2024, the preservation number is GDMCC No: 6528, and the preservation number is CGMCC NO: 6528. The drug-resistant cell strain MDA-MB-231 / CBP constructed in the invention can stably grow and pass in a carboplatin culture system with an action concentration of 20 [mu] M, has a carboplatin resistance index (RI) of 10.7, provides a basis for research of a carboplatin drug-resistant mechanism and development of related targets, and has a wide application prospect. The invention provides a drug-resistant cell model for researching the drug resistance mechanism of human breast cancer cells to carboplatin, searching an effective treatment method for overcoming human breast cancer drug resistance, preparing antitumor drugs, screening tumor drug resistance reversal drugs, guiding clinical medication and the like, and has important effects and good application prospects.

The invention relates to the field of cell engineering, in particular to application of Siglec7 in generation and / or release of exosomes in tumor cells. According to the invention, after a lentiviral vector of an overexpressed Siglec7 gene is constructed, tumor cells are transfected with the lentiviral vector, and finally the tumor cell strain stably expressing Siglec7 is obtained. Through extraction and detection, the cell strain obviously improves the yield of the exosome in unit cell and unit culture medium volume, and the yield of the exosome is obviously improved.

The invention discloses a method for directly inducing generation of organoids from IPS and application, and belongs to the field of cell engineering. The invention provides a method for directly and rapidly generating a target organ based on human IPS, and the lung organ, the placenta organ and the blood vessel organ are obtained. According to the method, the organoid represented by three germ layers is induced, so that the method is suitable for constructing various organoid. Compared with the prior art, the method has the advantages that step-by-step induction is not needed, one-step construction is successful, the operation is simple, the construction time is greatly shortened, the process is repeatable, long-term stable culture and passage are realized under in-vitro conditions, and the freezing survival rate is high; the method has the advantages that the method is simple in operation, the cultured organoid is good in homogeneity, RNA-seq results show that high expression of the blood vessel related genes appears in the organoid, and the maximum length of the organoid can reach 400 micrometers, so that the culture method can spontaneously induce expression of part of the blood vessel related genes to support growth of the organoid. The invention provides an important tool for researching pathogenesis of diseases and subsequent drug development.

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

The invention belongs to the technical field of biology, and relates to an anti-H6 subtype avian influenza virus hemagglutinin protein monoclonal antibody 1H1 and application thereof, cell engineering and antibody engineering technologies are utilized to obtain a hybridoma cell line for secreting a hemagglutinin monoclonal antibody, ascites is induced by mice of the same strain to prepare the anti-hemagglutinin protein monoclonal antibody 1H1, the anti-hemagglutinin protein monoclonal antibody 1H1 is identified as an IgG1, kappa type, and the anti-hemagglutinin protein monoclonal antibody 1H1 is identified as an anti-H6 subtype avian influenza virus hemagglutinin protein monoclonal antibody. And the application of the antibody is realized through technologies of affinity purification, immunization and the like. The monoclonal antibody has the advantages that the preparation method is simple and easy to implement, more importantly, the monoclonal antibody prepared by the method can have multiple purposes, an effective tool is provided for auxiliary diagnosis of H6 subtype avian influenza virus infection in a clinical sample, and the monoclonal antibody can be popularized and applied to multiple detection technologies and clinical and experimental research.

The invention belongs to the technical field of cytobiology and regenerative medicine, and particularly relates to a porcine-derived corneal limbus epithelial stem cell culture method and application, during culture, pretreatment is firstly carried out, then a culture dish is coated, cells are cultured and passaged after centrifugation, dryness identification is carried out through cell immunofluorescence color, and the porcine-derived corneal limbus epithelial stem cells are obtained. The stem cell engineering vector can be applied to damage repair of corneal diseases including corneal limbal stem cell deficiency, during repair, the stem cell engineering vector is compounded with a biological material to form the stem cell engineering vector, the stem cell engineering vector is implanted into a damaged part, the separation and culture method is simple in step, the cell obtaining amount is large, and the stem cell engineering vector is suitable for being separated from eyeballs of a large animal-pig; the corneal limbus epithelial stem cells cultured in vitro can form cell clusters called stem cell islands, the obtained corneal limbus epithelial stem cells have phenotypes of BMI1 + and delta Np63 alpha +, and CK14 + and ABCG < 2 + >, and a new way can be provided for cell transplantation based on experimental research and clinical research.

The invention relates to the technical field of test tube cleaning and disinfection, and discloses a device for cleaning and disinfecting test tubes after biological cell engineering cultivation, the device comprises a bottom frame, through a designed driving motor and a threaded rod, a placing table for placing test tube bodies can be driven to transversely slide; the placing table drives the test tube body to move into the disinfection bin, the flushing bin, the scrubbing bin and the drying bin, and finally returns to the initial position to be irradiated and disinfected by the disinfection lamp, multiple functions of disinfection, flushing, scrubbing, drying and the like are integrated, the cleaning and disinfection process of the test tube can be completed in a one-stop mode, time and space are saved, meanwhile, manual operation is effectively reduced, and the working efficiency is improved. The cleaning and disinfecting efficiency and consistency are improved, the time-consuming and labor-consuming operation that an operator needs to soak, brush, wash and disinfect the test tubes one by one is avoided, and the problem that the cleaning and disinfecting effects are uneven due to the difference between different operators is solved.

Intracellular delivery of a genetic construct to immune cells including: obtaining a deterministic mechanoporation (DMP) platform that includes a substrate having a surface and a plurality of capture sites, each said capture site having a boundary shape at the surface adapted and configured to support thereon a cell, and each said capture site having a bottom and including a sub-micron-scale projection extending from the bottom toward the surface of the substrate, wherein said projection is adapted and configured to penetrate a cell membrane and / or wall of the cell, and wherein the substrate has a plurality of aspiration vias situated at the bottom of the capture sites; introducing the cells to the surface in a liquid media; capturing the cells within the capture sites by applying a first hydrodynamic force; applying a second hydrodynamic force on the captured cell and locally rupturing the membrane and / or wall of the cell with the projection, introducing the genetic construct into the cells, and releasing the porated cells from the capture sites. Also disclosed are methods of chimeric antigen receptor (CAR) T cell adoptive immunotherapy and T cell receptor (TCR) therapy.

The invention relates to the technical field of gene engineering and cell engineering, in particular to an sgRNA combination for precisely editing a dairy cow BLG gene, a primer and application. The sgRNA combination comprises: sgRNA1, the sequence of which is as shown in SEQ ID NO. 1; the sequence of the sgRNA2 is as shown in SEQ ID NO. 2; the sequence of the sgRNA3 is as shown in SEQ ID NO. 3; and the sequence of the sgRNA4 is as shown in SEQ ID NO. 4. The BLG gene edited mammary epithelial cells and the BLG gene single base edited dairy cow can be successfully obtained, and the method has a wide application prospect in the aspect of dairy cow breeding.

The invention belongs to the technical field of cell engineering, and particularly relates to a construction method and application of a mammal rapamycin target protein self-activated mouse model and a mouse embryo fibroblast line. According to the invention, serine (Serine, S) at site 2215 of mTOR protein is mutated into tyrosine (Tyrosine, Y), and a transgenic mouse model with activation mutation of mTORS2215Y, namely a mouse model with self-activation of mammal rapamycin target protein, is constructed. A mouse embryo fibroblast line which can be immortalized is constructed by utilizing the transgenic mouse model, so that the related mechanism research of mTOR self-activation mutation can be carried out at the cellular level, and the screening of related therapeutic drugs can be carried out.

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.