233 results about "Cell engineering" patented technology

The present invention is in the field of CRISPR-Cas system for genome targeting. The present invention relates to new engineered Cas9 scaffolds and uses thereof. More particularly, the present invention relates to methods for genome targeting, cell engineering and therapeutic application. The present invention also relates to vectors, compositions and kits in which the new Cas9 scaffolds of the present invention are used.

The preparation process of porous polylactic acid microball includes the following steps: dissolving polylactic acid in solvent dichloromethane and adding bad solvent via mixing; dissolving PVA in deionized water to prepare solution, and under stirring, pouring the polylactic acid solution into the PVA solution; volatilizing organic solvent, filtering and washing to obtain the porous polylactic acid microball. The said process is simple and has mild condition, and the obtained porous polylactic acid microball may be used widely in controlled medicine releasing system, micro cell carrier in cell engineering and adhered microball rack in tissue engineering.

The invention discloses a method for preparing nano hydroxyapatite/polylactic acid composite microspheres, which uses nano hydroxyapatite (n-HA) and polylactic acid (PLA) as raw materials and adopts an ultrasonic blending composite process and an emulsification-solvent evaporation method to prepare the microspheres of a nano hydroxyapatite/polylactic acid composite material. The preparation method comprises the following steps: nano-crystallizing hydroxyapatite, proportionally preparing complex liquid, performing ultrasonic dissolving and blending, emulsifying the mixture into small liquid drops, reducing pressure to volatilize a solvent, performing refrigeration and filtration, performing washing, and performing freeze-drying to obtain the composite microspheres. Compared with like products, the composite microspheres have the characteristics that the method has simple and easy operation, the size of the microspheres is easy to control, and the prepared microspheres have large surface holes and specific area, and good mechanical property. The prepared nano hydroxyapatite/polylactic acid composite microspheres are mainly applied to microsphere bonding type brackets in bone tissue engineering and cell micro-carriers in cell engineering, and can be applied to conveying medicaments and bioactive molecules.

The invention belongs to the fields of genetic engineering and cell engineering, pertaining to a human T-lymphocytic series model for screening and reactivating latent infection HIV-1 compound and a preparation method thereof. The invention discloses a T lymphocyte used for screening and activating latent infection HIV-1 drug, and the T lymphocyte is a Jurkat stable strain that is infected by but not expresses HIV slow virus carrying reporter gene. The preservation number of the T lymphocyte is CCTCC NO.C200821. The invention also provides a preparation method for the T lymphocyte, including infecting human T-lymphocytic series Jurkat with the HIV-1 slow virus carrying EGFO reporter gene, and carrying out cell sorting and HIV integration test so as to obtain the clone with HIV integration while not expressing EGFP. The invention is the cell model for the virus preservation library to control drug screening.

The invention discloses a base station coverage and basic data inspection method and device. The method comprises the steps that an adjacent cell level switching report of a serving cell is collected, and a target cell with the maximum switching value is screened out; cell engineering parameter lists of the serving cell and the target cell are colleted; the interstation distance between the serving cell and the target cell is calculated according to the geographic position information of the serving cell and the geographic position information of the target cell, and the position relevance between the serving cell and the target cell is determined based on the interstation distance; the coverage relevance of the serving cell and the target cell is calculated according to the geographic position information and the coverage performance parameters of the serving cell and the geographic position information and the coverage performance parameters of the target cell; if the position relevance or the coverage relevance is zero, it is determined that actual configuration data of a first base station or/and a second base station are not matched with preset configuration data. The abnormal base station can be accurately positioned in time, the fault base station can be examined on the spot in a targeted mode, the inspection of regional and even the whole network base station coverage and basic data is achieved, the inspection difficulty and the checking workload are reduced, and the working efficiency is effectively improved.

The invention belongs to the field of cell engineering, in particular to a method for high-efficient expansion and cryopreservation of NK (Natural Killer) cells and an application of the method. According to the invention, fusion genes are subjected to gene recombination to an AAVS1 (Adeno-Associated Virus Integration Site 1) transposon through a plasmid, and the freezing/thawing step is introduced to the NK cell expansion step taking a modified K562 cell line as a basis in the intermediate stage of cell expansion; finally, the NK cells are expanded and saved. A K562-mbIL15-41BBL-mbIL21 cell line developed by the integration technology of site specific genes is used, and the activity, purity and cytotoxicity of the NK cells are high to realize high expansion efficiency of the NK cells to improve NK expansion. The method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells and the application of the method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells disclosed by the invention have great market prospects and economic values.

The invention discloses a method for preparing and regenerating a phomopasis asparagi protoplast, which relates to the technical field of fungus protoplast preparation and regeneration in cell engineering. The method comprises the following detailed operation steps of: (1) preparing a phomopasis asparagi conidium suspension; (2) preparing a fresh mycelium; (3) performing enzymolysis on mycelium cell walls; (4) separating the protoplast; and (5) regenerating the protoplast. The method is easy to operate, low in requirements on equipment and short in enzymolysis time and regeneration period, and effectively enhances the efficiency of preparation and regeneration of the protoplast.

The invention discloses a preparation method of a hydroxy apatite / polylactic acid / chitosan composite microballoon. The hydroxy apatite / polylactic acid / chitosan composite microballoon is prepared by using hydroxy apatite HA, polylactic acid PLA and chitosan CS as raw materials and employing manufacturing principles of a solution blending method and an emulsion method. The method comprises steps of: dropping a prepared apatite / polylactic acid / chitosan acetate acid gracial solution into a stirred liquid paraffin / petroleum ether mixed liquor ; carrying out stewing; removing the liquid paraffin / petroleum ether mixed liquor; washing; carrying out stewing; and refrigerating drying to obtain the composite microballoon product. The hydroxy apatite / polylactic acid / chitosan composite microballoon of the invention compounds three materials with different performances to solve problems of strength, toughness and biocompatibility of the material; and the hydroxy apatite / polylactic acid / chitosan composite microballoon is mainly used in bone tissue restoration material, cell engineering and transmission of medicament and bioactive molecule.

The invention discloses a cell separating method and special cell separating medium. The special cell separating medium gains is solvent gained by adding surface active agent into general cell separating medium. The cell separating method includes the following steps: mixing the surface active agent and current cell separating medium in given volumetric proportion; centrifuging to gain special cell separating medium; adding cell separating sample into the special cell separating medium; and centrifuging; colleting cloud layer cell and washing to gain purified cell. The invention greatly shortens sample adding and cell separating time, simplifies operation. And it is fit for large-scale separating. Its effect is stable and reliable. And high purity goal cell can be gained. The separating method is simple, and has strong operability. The invention has high practical application value and broad market prospect in cell biology, functional genome, and cell engineering fields.

The invention provides a recombinant lentiviral expression vector, and belongs to the technical field of cell engineering. The recombinant lentiviral expression vector comprises a 21mb gene, a 41BBL gene and an MICA gene which are sequentially connected in series; and the 21mb gene is obtained by sequentially connecting a signal peptide gene, an IL21 gene, a CD8 Hinge gene and a CD8 transmembranegene in series. After the provided recombinant lentiviral expression vector transfects a host cell, the host cell expresses the MICA and the 41BBL, and the IL21 is fused and expressed (but not secreted in a culture environment) on a cytomembrane of the host cell. After the provided recombinant lentiviral expression vector transfects the host cell, an obtained reconstitution cell promotes the highexpression of the NKG2D, avoids excessive activation of an NK cell and prolongs the proliferation time of the NK cell in the process of induction culture of the NK cell.

The invention relates to the field of cell engineering and discloses mesenchymal stem cells, as well as a preparation method and an application thereof. The method comprises the following steps of cutting an umbilical cord into small pieces after removing an umbilical artery and umbilical veins, digesting with an alpha MEM (minimum essential medium) culture medium containing 0.1% of type IV collagenase and a complete medium, then centrifugating cell suspension obtained after digestion, removing supernatant liquid, then washing, further performing resuspension with the complete medium, transferring suspension after resuspension into a culture bottle coated by a wall-adhered matrix for culture till the emergence of shuttle-shaped wall-adhered cells, washing, and replacing the new complete medium for continuing the culture so as to get the shuttle-shaped wall-adhered cells after the culture, namely the mesenchymal stem cells. According to the preparation method disclosed by the invention, the culture medium without serum or foreign protein is used for preparation, a recombinase with a single component is used for passage, and the prepared mesenchymal stem cells are smaller in diameter, have no sensibiligen and can improve the vitality of the cells and increase the safety in regeneration treatment of a patient.

The invention discloses a method for wheat tissue culture, which comprises the following steps: firstly, performing induction culture on a wheat germ to obtain an embryogenic callus; and secondly, performing differentiation culture on the embryogenic callus to obtain a wheat regenerated plantlet, wherein the method of the induction culture comprises the steps of successively performing induction cultures on the wheat germ with a culture medium containing auxin, a culture medium without the auxin, and the culture medium containing the auxin again to obtain the embryogenic callus. The method for the wheat tissue culture further improves the regeneration frequency of immature embryos and mature embryos of wheat, weakens the barriers caused by the conditions such as genotypes, physiological states and the like to the regeneration, provides a technical support for the cell engineering breeding, the genetic engineering breeding and the functional genomics research of the wheat, and has great significance for the biotechnology breeding and the functional genomics research of the wheat.

This invention discloses monoclonal antibody against anti-LCDV immunoglobulin of Paralichthys olivaceus, which is excreted by hybridoma JF-lgM-H (CCTCC-C200631). The method comprises: immunizing Paralichthys olivaceus with LCDV inactivated by formalin to prepare antiserum, purifying Paralichthys olivaceus immunoglobulin, immunizing Balb/c mice as antigen, preparing hybridoma cells by cell engineering method, and screening the monoclonal antibody by immunoassay. Indirect ELISA and indirect immunofluorescent antibody assay show that this monoclonal antibody is located on the heavy chain (7-80 kDa) of the anti-LCDV immunoglobulin. The monoclonal antibody can be used for preparing reagents for detecting LCDV infection in early stage, and evaluating the immune effects of LCDV vaccine inactivated by formalin.

The invention belongs to the technical field of cell engineering, and specifically relates to a method for obtaining Lolium L. embryogenic callus and maintaining regeneration capacity of the callus. According to the present invention, the material for callus induction is the mature seed embryos of the Lolium L.; the mature embryos are adopted to induce the callus, after the mature embryos are cultured for 1 month, 4 secondary cultures are performed, wherein the time of the 4 secondary cultures is 2 months, and the secondary culture is performed every 2 weeks; a differentiation regeneration culture treatment is performed on the callus, the differentiated plant is continuously subjected to the secondary culture, and the callus incapable of differentiation is discarded; the stipe meristem area of the secondary culture plant after regenerating is subjected to the second callus induction, wherein the culture time is 1 month; the callus is subjected to the secondary culture for 2 months, wherein the culture media is changed every two weeks; then the differentiation regeneration culture treatment is performed on the callus, wherein the callus incapable of differentiation is discarded; a long-term secondary culture is performed on the regenerated plant, when the callus is required to carry out the relevant research, the stipe meristem area of the plant is subjected to induction and secondary culture to obtain the required callus, wherein the regenerated plant is subjected to the second differentiation. According to the present invention, the operation is simple and feasible, and the stable and high-frequency differentiation regeneration efficiency can be obtained.

The method of separating and purifying chickení»s spermatospore relates to trnasgene, rare species preservation, tissue and cell engineering, and other fields. The present invention includes first separating chickení»s spermatospore and subsequent Percoll separation and adherent purification. The present invention provides proper chickení»s spermatospore separating and purifying method by means of separating chickení»s spermatospore from chickení»s testis tissue, and Percoll gradient and adherent purification, spermatogenic epithelium cell suspension with great testis cell number and high survival rate of testis cell is prepared while ensuring elimination of other somatic cell components.

The invention discloses a nanometer artificial seed of Dendrobium officinale and a manufacturing method thereof. The manufacturing method comprises the following steps: induction and propagation of protocorm of Dendrobium officinale; and manufacturing and germination of artificial seeds. According to the invention, nanometer technology and cell engineering technology are combined together to prepare composite endosperm and artificial seed coat of the artificial seed of Dendrobium officinale, so permeability and water retention performance of traditional embedding are improved and the growth of the protocorm is promoted; thus, the germination rate and the seedling rate of the artificial seeds are increased, and technical support is provided for practicability of germplasm breeding and industrialization development of Dendrobium officinale.

The invention discloses a continuous pouring culture device of plant cell stirring typed biological reactor in the cell engineering technical domain, which comprises the following parts: one-grade sediment device, two-grade sediment device, three-grade sediment device, liquor storage bottle, sampling device, on-line monitor system, gas supplying device, sediment device in connection with the reacting tank on the magnetic stirrer, wherein the waste liquor bottle connects the sediment device. The invention is a group of three-gravity inner-outer cylinder biological reactor of sediment cell interception device to provide reference to culture large scale of plant cell, which improves the separating efficiency of the cell and culture liquid of reactor to reduce the probability of pollution.

The invention discloses a fabrication method of seagrass seedling protoplast, comprising the following steps: temporarily raise the seagrass seedlings in double-anti sea water, select the seedlings that just sprout and sequentially put them into alcohol with the concentration of 75 percent, sodium hypochlorite solution with the concentration of 1 percent and potassium iodide solution with the concentration of 1.5 percent for disinfection, then wash, soak and restore the seedlings by sterile seawater. Then, the seagrass seedlings are shredded and placed into centrifugal tube with enzyme solution for enzymolysis. After the enzymolysis, glucose liquid is added before filtration. Wash the seedlings thrice with hypertonic sea water and obtain the precipitate after centrifugation. Add medium to the precipitate and obtain the protoplast through centrifugation. The invention can avoid pollution and obtain sterile seagrass seedling protoplast and has laid a solid foundation for the protoplast fusion, genetic transformation, plant regeneration as well as new breeds breeding. Moreover, the invention can also provide technical reserves for seagrass propagation of cell engineering, rapid propagation of fine lines seagrass and improving seagrass genetic engineering.

The present invention is process of inducing indefinite bud and somatic embryo of North America redwood leaf and regenerating plant, and belongs to the field of forestry cell engineering breeding technology. The technological scheme includes inoculating stem apex of North America redwood in MS culture medium for secondary culture, taking 28-32 day and 30-40 day cultured test tube plantlet and making the far axial plane contact with the culture medium to dark culture for 30 days, transferring to fresh culture medium to culture for other 20 days, and transferring for light culture. In SH culture medium containing BA 0.5 mg/L+KT 0.2 mg/L+IBA 0.2 mg/L and BA 0.5 mg/L+IBA 0.5 mg/L, indefinite bud and somatic embryo are induced successfully. The present invention makes it possible to realize the large scale, short period and low cost industrial production of North America redwood nursery stock, and has important cytology significance.

The invention provides a preparation method of a human dendritic cell tumour vaccine and belongs to the technical field of cell engineering. The human dendritic cell tumour vaccine is characterized in that a DC maturity acceleration agent is added in a process that the dendritic cell is matured from being unmatured, and the DC maturity acceleration agent comprises OK-432 and a gp100 combined DC maturity acceleration agent. By adopting the human dendritic cell tumour vaccine, cancer patients can be treated or high risk of people suffering from cancer can be avoided. The human dendritic cell tumour vaccine prepared by adopting the preparation method provided by the invention has the advantages that DC purity and maturity of the human dendritic cell tumour vaccine are greatly improved, fusion of DC and a tumour antigen is promoted, and treatment of cancer can be facilitated; meanwhile, a preparation technology is simple, and cost is low.

The invention discloses an application of tri-methylation of histone H3lysine 4( H3K4me3) to granulosa cells of pigs, and belongs to the technical field of cell engineering and genetic engineering. H3K4me3 is used as a penetration point, and a cell biology method is used for researching the influence of the H3K4me3 to the functions of the granulosa cells of pigs. The technical scheme is detailed in design, and reliable in results. In order to confirm the influence of H3K4me3 to the functions of the granulosa cells of the pigs, verification is performed from multiple levels and multiple angles,and verification is performed from the cell level and the protein level. The invention explains that the H3K4me3 has influence on the functions of the granulosa cells of the pigs, the H3K4me3 can promote proliferation of the granulosa cells of the pigs, can restrain the apoptosis of the granulosa cells of the pigs, and can reduce the proportion of the granulosa cells of the pigs, blocked in the GO/G1 period. The application has a good application value of research of histone methylation on an influence mechanism on the development of granulosa cells and the reproductive performance of sows.

The invention belongs to the technical field of cell engineering, and particularly relates to a cytomechanic-electrical joint loading and analyzing device in a biomedical experimental instrument. The invention discloses a device for cell composite force-electric load measurement. The device comprises a mechanical drive system, a mechanical drive control system, an electrode probe, a cell placement platform, an electric signal generation system and a piezoelectric measurement system. The invention also discloses a device for cell composite force-electric load measurement. The device for cell composite force-electric load measurement comprises a mechanical drive system, a mechanical drive control system, an electrode probe, a cell placement platform, an electric signal measuring system and a piezoelectric measurement system. The devices disclosed by the invention are applied to an application prospect for cytomechanic-electrical load measurement on cells.

The invention relates to the domain of cell engineering, in particular to a method for rejuvenating and detoxifying an edible fungal strain by using protoplast regeneration technology. The method comprises the following steps: selecting parents, preparing protoplast, regenerating the protoplast, and establishing an evaluation standard for rejuvenating and detoxifying the edible fungal strain. The step of preparing the protoplast comprises the step of selecting steady infiltration agent solute, steady infiltration agent concentration, degrading enzyme, enzymolysis time and enzymolysis temperature; and the step of regenerating the protoplast comprises the step of selecting a pre-culture medium and a culture method. The method has the advantages that the effects of removing viable bacteria in the edible fungal and inhibiting factor accumulation are achieved by obtaining the protoplast so as to fulfill the purposes of rejuvenating and detoxifying the edible fungal and recover good properties of the edible fungal. For example, the comprehensive properties of oyster mushroom 89 and pleurotus ferulae 10 are remarkably superior to those of a contrast variety in the aspects of yield, adaptability, commercial property, impure fungal resistance and the like, and the remarkable economic benefit is produced.

The external cultivating method of primordial bird germ cell relates to transgene, animal cloning, rare animal preservation, tissue and cell engineering and other science fields. After it is inoculated to the CEF raising layer, the primordial bird germ cell is cultivated in liquid high-sugar DMEM cultivation medium with added calf serum in 10 úÑ, chicken serum in 2 úÑ, L-glutamine in 2mmol/L, sodium pyrovate in 1 mmol/L, beta ¿Cmercaptoethanol in 5.5í‡10-5 mol/L, optional amino acid 10 microliter/ml, hSCF in 15-20 ng/ml, mLIF in 10 U/ml, bFGF in 10-15 ng/ml, hIL-11 in 0.04 ng/ml, IGF in 10 ng/ml, and gentamycin sulfate in 100 U/ml. The present invention has simple operation and high repeatability, and lays the foundation of the relevant research.