37 results about "Cells/microL" patented technology

The invention discloses micro ribonucleic acids (microRNA, miRNA) carried by cell microparticles (Microparticle, MP), a method for preparing the same, and application thereof in the technical field of biotechnological pharmacy. The invention provides a combination of the micro ribonucleic acids for evaluating the physiological and / or pathological states of a participant, and the combination contains all the micro ribonucleic acids which exist stably in serum / plasma particles of the participant and are detectable. At the same time, the invention provides an experimental method for preparing the cell microparticles containing specific micro ribonucleic acids and using the cell microparticles to perform gene-level regulation and control as well as modification on other cells and tissues. The combination and the method can be used for detecting and treating various diseases, including the aspects of the diagnosis and the differential diagnosis of various tumors, various acute and chronic infectious diseases and other acute and chronic diseases, the prediction and the curative effect evaluation of the occurrences of disease complications and the recurrences of malignant diseases, as well as the active ingredient screening, the efficacy evaluation and the judicial authentication of drugs, the detection of prohibited drugs and the like; besides, the combination and the method have the advantages of wide detection pedigree, high sensitivity, low detection cost, convenient available material, easy storage of samples and the like.

The present invention relates a method for the enumeration of eukaryotic cell micronuclei, while simultaneously acquiring cytotoxicity and mode of action information. The method utilizes differential labeling of chromatin from dead and dying cells to distinguish the chromatin from micronuclei, nuclei, and metaphase chromosomes, and differential labeling of metaphase events to provide additional information regarding cytotoxicity and genotoxic modes of action. Counting of micronuclei events relative to the number of nuclei and quantifying perturbations to the proportion of metaphase events can be used to assess the DNA-damaging potential of a chemical agent, the DNA-damaging potential of a physical agent, the effects of an agent which can modify endogenously-induced DNA damage, the effects of an agent which can modify exogenously-induced DNA damage, and genotoxic mode of action.

The invention relates to a preparation method and application of an anti-tumor vaccine based on cell microvesicles and aims to effectively solve problems in preparation of anti-tumor vaccines high inyield, good in universality, strong in killing effect on tumor cells and capable of controlling tumors by improving tumor microenvironments, repairing an immune system and enhancing anti-tumor immuneresponse of organisms. The preparation method comprises the following steps: preparing tumor cell microvesicles loaded with immunomodulators and connecting the surfaces of the microvesicles with adjuvant-loaded liposome to stably form the anti-tumor vaccine. The prepared anti-tumor vaccine is high in yield, good in universality, strong in killing effect on tumor cells and capable of improving tumor microenvironments, repairing an immune system and enhancing anti-tumor immune response of organisms; the method is simple in process and high in efficiency; and the prepared microvesicles are derived from cells, are sufficient in quantity and wide in source, are easy for large-scale production, improve the bioavailability of adjuvants, are effectively applicable to preparation of anti-tumor vaccines based on the cell microvesicles, and can be applied to prevention and treatment of different types of tumors.

The invention discloses a method for preparing virus of porcine reproductive and respiratory syndrome on a large scale. In the method, the virus of the porcine reproductive and respiratory syndrome is prepared in a cell microcarrier suspension culture system by a bioreactor. The method comprises the following steps of: inoculating host cells for preparing the virus to a carrier tank containing culture solution and a microcarrier, and mixing the cells and the microcarrier uniformly to ensure that the cells are attached to the microcarrier; providing sufficient nutrients and appropriate gas environment for the cells under the appropriate culture environment to ensure that the cells are grown until the cells are in an amount which are 10 to 20 times of the inoculation concentration on the microcarrier; preparing virus suspension from the virus of the porcine reproductive and respiratory syndrome by using cell maintenance culture solution to ensure that the suspension is adsorbed to the cells; culturing the virus under the appropriate culture environment; culturing continuously for 2 to 3 days to obtain virus solution; and after the virus solution passes inspection, performing freeze thawing on the virus solution twice at the temperature of -20 DEG C, and inactivating and purifying to prepare an inactivated vaccine of the porcine reproductive and respiratory syndrome or adding a freeze-drying protective agent for freeze drying to prepare a live vaccine of the porcine reproductive and respiratory syndrome. The method has large production scale, high yield of single batch and low production cost.

The invention relates to a serum-free medium adapting to PK-15 all-suspended growth, a preparation method of serum-free medium and an all-suspended domestication method applied to a PK-15 cell. The serum-free medium comprises amino acid, Tween, linoleic acid, vitamin E, myristic acid, stearic acid, vitamin A, beta-mercaptoethanol, sodium chloride, glucose, cadmium chloride, linoleic acid, cholinechloride, inositol, ammonium metavanadate and the like. The provided serum-free medium can realize all-suspended growth of the PK-15 cell, so that the cost and digestion of a microcarrier are omitted,and the technical problem that the PK-15 cell microcarrier is difficult to digest and amplify in the suspended culture process is solved.

The present invention provides a cell-microsphere with an eluted superparamagnetic gel coating and a preparation method thereof. The cell-microsphere comprises a gel microsphere and a super-paramagnetic sphere coating the gel microsphere. Magnetic gel coating, the gel microspheres contain cells, the superparamagnetic gel coating contains superparamagnetic particles, the superparamagnetic gel coating can be removed by elution, and the superparamagnetic gel The gel coat and gel beads cannot be removed by simultaneous elution. The present invention also provides the application of the above-mentioned cell-microspheres in the co-culture of cells containing cell-gel microspheres for clinical implantation and repair and the separation in the later stage of the co-culture. The invention can solve the problems that the existing microcarrier cannot realize the removal of magnetic material and the separation of different cells in the co-culture system after the co-culture, and cannot be used for clinical implantation and repair.

The invention discloses a preparation method of immobilized cells. The method comprises the following steps: a pectin solution is added to a suspension containing to-be-immobilized cells, a calcium chloride and chitosan solution is added, and a pectin-containing cell microspherical suspension is obtained. According to the method, the cell activity can be maintained, nutrients of the cells are allowed to enter pellets, regular growth and division of the cells are kept, the pellets can be secreted by metabolites of the cells, and accordingly, related recombinant proteins are separated from a culture medium and purified. The invention further discloses applications of the preparation method to the immobilized cell engineering and drug preparation.

The invention discloses core-shell-structured microspheres for monitoring mechanical properties and contraction frequency of myocytes as well as application thereof, and belongs to the technical fieldof biomedicine. At present, studies on cell contraction force are mainly based on two-dimensional cell models, so that in vivo environment of myocytes cannot be completely simulated. The invention provides three-dimensional microspheres with core-shell structure. Each of the three-dimensional microspheres with core-shell structure comprises a core layer and a shell layer; the core layer is a sphere formed by performing solidification on a first gel material; the shell layer is a shell composed of cells and a second gel material, and wraps the core layer; and the core layer deforms along withpulsation of the cells in the shell layer, so that, distribution of forces produced by the shell-layer cells on the core layer can be obtained by observing the deformation of the core layer. Comparedwith two-dimensional cell culture, the three-dimensional cell culture system focuses on cell-to-cell contact and cell-to-matrix contact, so that, growth environment of organisms can be better simulated; and thus, the three-dimensional microspheres with core-shell structure can be applied in disease research cell models and drug screening models.

The invention provides a cell microcarrier matrix. The raw materials include methacrylated gelatin and oyster shell powder. According to the mass ratio, methacrylated gelatin: oyster shell powder=1-10:0.5-2. Methacrylated gelatin has high water content, similar composition to extracellular matrix, and has good surface activity, so it can be well compatible with cells and active factors. The carrier matrix wraps the cells in it, which can provide a three-dimensional support for the cells, and can also prevent the cells from being damaged during injection and transplantation. The compound use of methacrylated gelatin and oyster shell powder can effectively reduce the swelling rate of the cell microcarriers, so that the cell microcarriers have better mechanical strength.

The present invention relates to a serum-free medium adapted to PK-15 full-suspension growth, a preparation method thereof and a full-suspension acclimation method applied to pK-15 cells. The medium contains amino acids, Tween, linoleic acid, vitamin E, Myristic acid, stearic acid, vitamin A, β‑mercaptoethanol, sodium chloride, glucose, cadmium chloride, linoleic acid, choline chloride, inositol, and ammonium metavanadate. The serum-free medium provided by the present invention can realize the full suspension growth of PK‑15 cells, thereby omitting the cost of microcarriers and the digestion process, and solving the technical problem that it is difficult to digest and enlarge the PK‑15 cell microcarrier suspension culture process.

The invention discloses a tumor cell microsatellite unstable state detection system, which relates to quantitative fluorescent PCR amplification technology and capillary electrophoresis detection technology. This system can amplify the six single nucleotide repeat sites of NR‑21, NR‑24, NR‑27, MONO‑27, BAT‑25, BAT‑26, and simultaneously amplify Braf V600E, UGT1A1*6, UGT1A1*28 loci were typed. On the basis of determining the status of MSI, the detection results can realize the screening of Lynch syndrome and provide reference for the use of irinotecan drugs. The invention also provides a detection kit designed accordingly.

The present invention provides a cell reactor microcarrier cell harvesting and re-inoculation amplification method, comprising step 101: filtering and collecting the microcarrier cells; step 102: removing the medium for buffer balance; step 103: trypsinization; step 104: cell Microcarrier isolation, harvesting cells only or adding next-level microcarriers to inoculate scale-up. The invention solves the problems of the collection of production-scale microcarriers, the controllable digestion of cells, the separation of the harvested microcarriers of cells, and the amplification of spheres. The method of the invention is suitable for the preparation and production of adherent cells, and is applied to the pilot scale and production of vaccines, recombinant proteins and biological enzymes, and the method is simple and economical.

The invention provides an enhanced interception device and method for perfusion culture of cells with microcarriers, wherein a layered funnel is arranged inside the device body, and the bottom of the layered funnel is a settling chamber, and a culture medium-microcarrier inlet is opened on the settling chamber, and the sedimentation The bottom of the cavity is connected with a connecting pipe, the bottom of the connecting pipe is the medium-microcarrier return port, the top of the layered funnel is the outlet of the medium, and there is an observation window at the layered funnel on one side of the device body; the advantage is that the present invention has solid-liquid settlement The function of separating and controlling the liquid level can improve the perfusion volume and interception efficiency of cell microcarrier culture, solve the problem of microcarrier loss due to not easy sedimentation when the liquid feeding speed is too high, and cause less damage to cells during the process of microcarrier retention.

The invention relates to a cell microcapsule and a preparation method thereof, a cell microcapsule loaded with cancer resisting medicines, and a preparation method and application of the cell microcapsule loaded with cancer resisting medicines. The preparation method of the cell microcapsule comprises the following steps of (1) culturing immunocyte, and taking cell supernatant; (2) centrifuging the cell supernatant obtained in the step (1) to remove cells and cell fragments, and taking supernatant; (3) centrifuging the supernatant obtained in the step (2) to remove apoptotic bodies, and takingsupernatant, wherein the centrifuging condition includes that the centrifugal force is 2000-3000g, and the centrifuging time is 40-100min; and (4) centrifuging the supernatant obtained in the step (3), and taking precipitate. The preparation method is simple to operate, the cell microcapsule which is good in bioactivity, complete in structure, high in purity and uniform in particle diameter can be obtained; the obtained cell microcapsule can load liposoluble cancer resisting medicines through simple incubation, and the cell microcapsule is good in effects, good in targeting properties and lowin toxic and side effects when being used for preventing or treating cancer diseases.

The invention discloses cleavable cell microcapsules, a preparation method and a cell culture method thereof. The lysable cell microcapsules of the present invention include a capsule wall and cells wrapped by the capsule wall and culture fluid. The capsule wall can create an environment for long-term stable proliferation of cells. The cells grow adherently to meet the needs of cell morphology observation. The cell microcapsule of the present invention is particularly suitable for cell experiments in space environment.

A CRISPR / CasRx RNA editing system targeted inflammatory factor-based cell microvesicle is obtained by expression and secretion of engineering cells transfecting target plasmids, the cell microvesicle contains CRISPR / CasRx editing enzyme guided by an exosome signal peptide and sgRNA of the targeted inflammatory factor, and CRISPR / CasRx editing system targeted inflammatory factor-based cell microvesicle is obtained by expression and secretion of the engineering cells transfecting target plasmids under the guidance of a guide RNA sequence sgRNA of the targeted inflammatory factor. The mRNA of the inflammatory factor is specifically bound and degraded, and the mRNA level of the specific inflammatory factor in the target cell is reduced. The cell microvesicles have good fusion with host cells and low immunogenicity, and are used for treating acute inflammatory factor storm related diseases, such as acute respiratory distress syndrome (ARDS), cytokine release syndrome (CRS), septicemia and the like.

The present invention relates to a method for the in vitro production of mammalian neurons expressing 6 subtypes (2N4R, 1N4R, 0N4R, 2N3R, 1N3R, 0N3R) of Tau protein, comprising a neuronal differentiation step in which the cell microcompartment is cultured for a period of 5 to 100 weeks, the neuronal differentiation step is carried out in a bioreactor, each cell microcompartment comprising a hollow hydrogel capsule surrounding post-mitotic neuronal cells and extracellular matrix, the cell microcompartments remaining suspended in a housing of the bioreactor containing a neuronal differentiation medium.