599 results about "Cell culture" patented technology

The invention discloses an anti-human hepatitis B e antigen monoclonal antibody and application thereof, and relates to the technical field of hepatitis B detection. In a heavy chain variable region of the monoclonal antibody, an amino acid sequence of a complementarity determining region CDR1-3 is as shown in SEQ ID NO.1-3 or has at least 95% of homology with the sequence as shown in SEQ ID NO.1-3; in the light chain variable region, amino acid sequences of complementary determining regions CDR1 and CDR3 are shown as SEQ ID NO.4 and SEQ ID NO.5 or have at least 95% of homology with the sequences shown as SEQ ID NO.4 and SEQ ID NO.5, and an amino acid sequence of a complementary determining region CDR2 is FAS. The monoclonal antibody provided by the invention can well recognize a natural hepatitis B e antigen, and the titer of a supernatant cultured by a B cell can reach 1: 10000, so that the monoclonal antibody can be completely applied to detection research of the human hepatitis B e antigen. The chemiluminescence reagent prepared by using the monoclonal antibody has the advantages of high specificity, strong anti-interference capability, high detection sensitivity, good stability and the like, almost has no leak detection, and can replace an imported reagent.

The invention discloses a serum-free culture system and culture method of NK cells. The invention firstly discloses a serum-free culture system of the NK cells, wherein the serum-free culture system comprises a coating solution, an amplification culture medium, an activation culture medium and autologous plasma; the invention further discloses a culture method for NK cells by using the serum-free culture system. The method for culturing the NK cells by using the serum-free culture system disclosed by the invention reduces the culture cost of the NK cells, can simply, quickly, efficiently and safely obtain a large number of NK cells with high activity, high purity and strong killing ability in vitro, provides a better cell source for clinical application of the NK cells, and promotes further development of an NK cell treatment technology.

The invention belongs to the technical field of cosmetics, and discloses a wrinkle-removing and anti-aging composition containing recombinant collagen as well as a preparation method and application of the wrinkle-removing and anti-aging composition. The composition is prepared from the following components: 20.4 to 35.6 percent of glycerol, 1.1 to 3.5 percent of 1, 2-hexanediol, 8.2 to 15.4 percent of apple fruit cell culture extract, 1.6 to 3.2 percent of inositol, 0.2 to 1.0 percent of elastin, 0.1 to 1.7 percent of hydroxyproline, 0.1 to 1.0 percent of sodium polyglutamate, 0.1 to 1.0 percent of sodium hyaluronate, 0.01 to 0.1 percent of acetyl hexapeptide-8, 0.01 to 0.1 percent of fibronectin, 1.72 to 3.21 percent of soluble collagen and the balance of deionized water. The amino acid sequence of the soluble collagen is as shown in SEQ ID NO. 1. The skin care product prepared by adding the anti-wrinkle and anti-aging composition is good in safety and has the effects of resisting wrinkles, improving skin elasticity and resisting aging.

The invention relates to the technical field of stem cell culture, and one embodiment of the invention provides a stem cell differentiation culture device which comprises a culture box and further comprises an air inlet pipe, a main fan, a fixed box, a conversion cylinder, a filter cylinder, an auxiliary fan, a discharge box and a cleaning mechanism, the culture box is communicated with a temperature adjusting system through the air inlet pipe, the fixed box is communicated with the air inlet pipe, and the air inlet pipe is communicated with the main fan. The conversion cylinder is rotatably arranged in the fixed box, the interior of the conversion cylinder is divided into two conversion cavities, the filter cylinder is fixedly connected to the interior of the conversion cavities and communicates with the air inlet, the auxiliary fan is installed in the filter cylinder, the discharge box communicates with the fixed box through the discharge port, and the discharge box communicates with a discharge pipe. The cleaning mechanism is used for being matched with the auxiliary fan to clean the inner wall of the filter cartridge. By means of the technical scheme, the technical problem that dust accumulates on a filter screen in a ventilation system in the prior art, and regular disassembly and cleaning are needed is solved.

This invention discloses a monoclonal antibody against human hepatitis B e antigen and its application, relating to the field of hepatitis B detection technology. In the heavy chain variable region of the monoclonal antibody, the amino acid sequences of the complementarity-determining regions (CDR1-3) are as shown in SEQ ID NO. 1-3 or have at least 95% homology with the sequences shown in SEQ ID NO. 1-3; in the light chain variable region, the amino acid sequences of the complementarity-determining regions (CDR1 and CDR3) are as shown in SEQ ID NO. 4 and 5 or have at least 95% homology with the sequences shown in SEQ ID NO. 4 and 5, and the amino acid sequence of the complementarity-determining region (CDR2) is FAS. The monoclonal antibody provided by this invention can effectively recognize natural hepatitis B e antigen, and the titer of the supernatant from B cell culture can reach 1:10000, making it fully applicable to the detection and research of human hepatitis B e antigen. The chemiluminescent reagent prepared using the monoclonal antibody described in this invention has advantages such as high specificity, strong anti-interference ability, high detection sensitivity, and good stability, with almost no missed detections, and can replace imported reagents.

The invention relates to the technical field of synthetic biology, metabolic engineering and cell culture. The present invention provides a cell for the production of a negatively charged, preferably sialylated, oligosaccharide wherein the cell is genetically engineered to have or express, preferably overexpress, a hydrolytic UDP-N-acetyl-D-glucosamine-2-epimerase. The invention further provides the use of said cells in culture or incubation. Also described are methods of producing negatively charged, preferably sialylated oligosaccharides using the cells, and purification of the negatively charged, preferably sialylated oligosaccharides.

A method for constructing a microfluidic bone organoid-on-chip can continuously observe the impacts of different external factors on a bone tissue structure based on the “ternary regulation theory” of bone angiogenesis-bone resorption-osteogenesis coupling. The method includes: (1) building a visual three-dimensional cell cultivation platform to continuously observe impacts of different external factors on a ternary regulation theory based on angiogenesis of osteoclast precursors, osteoblasts and osteoclasts, such that a user is allowed to directly observe interactions among various kinds of cells; (2) designing microfluidic channels in a chip to allow a reagent involved to present a concentration gradient distribution in the chip; and (3) adding three bioinks including osteoblasts, osteoclasts and vascular endothelial cells respectively, so as to reflect the three cell components individually, in pairs, and as a whole in the chip. An apparatus for constructing a microfluidic bone organoid-on-chip is further provided.

The invention discloses a stem cell culture image identification and detection method, and relates to the technical field of stem cell image identification, and the method comprises the following steps: carrying out the graying processing of a stem cell image, obtaining a stem cell gray-scale image, carrying out the binarization processing of the stem cell gray-scale image, obtaining a stem cell binarization image, and obtaining a real-time complete contour based on the stem cell binarization image; acquiring a real-time length-width ratio; obtaining a range of a historical length-width ratio based on the first number of historical mesenchymal stem cells; the qualified real-time length-width ratio is obtained based on the range of the real-time length-width ratio and the historical length-width ratio, and the method is used for solving the problems that in an existing mesenchymal stem cell number recognition technology, the effective number of mesenchymal stem cells in the culture process cannot be accurately extracted, and consequently the accuracy and efficiency of recognizing the number of the mesenchymal stem cells are low; under the condition, the culture quality of the mesenchymal stem cells cannot be effectively judged.

The invention discloses a method for preparing human metapneumovirus nucleocapsid protein based on an insect-baculovirus system. Comprising the following steps: 1) constructing a recombinant vector with a human metapneumovirus nucleocapsid protein coding sequence: sequentially connecting a sequence for promoting secretory expression, the human metapneumovirus nucleocapsid protein coding sequence and a tag sequence from 5'to 3 'ends in the recombinant vector; 2) converting the recombinant vector with the human metapneumovirus nucleocapsid protein coding sequence constructed in the step 1) into a competent cell containing a baculovirus genome plasmid Bacmid to obtain a recombinant baculovirus genome plasmid; and 3) transfecting insect cells with the recombinant baculovirus genome plasmid obtained in the step 2), culturing and purifying to obtain the human metapneumovirus nucleocapsid protein. The invention provides the preparation method of the human metapneumovirus nucleocapsid protein, and a material support is provided for subsequent further research.

The present invention relates to a cell biomass (e.g. food product or pet food product) or its components prepared from cultivated metazoan cells, method of its preparation and a system for its preparation.

The invention provides bovine ephemeral fever virus positive serum as well as a preparation method and application thereof. The preparation method comprises the following steps: culturing bovine ephemeral fever virus by using MDBK suspension cells, inactivating by using beta-propiolactone, concentrating by using a 500kd hollow fiber column to obtain an antigen solution, and emulsifying the antigen solution and 61VG adjuvant according to a mass ratio of 1: 1.5 to prepare the vaccine; carrying out primary immunization and secondary immunization on the receptor, collecting blood of which the serum neutralizing antibody titer is greater than 1: 4096 14-21 days after the secondary immunization, and separating to obtain positive serum. The positive serum can be used for specific test, identification test and the like of the bovine ephemeral fever virus, can also be used for preparing related reagents, kits and standard substances, and provides efficient and normative technical support for prevention and control of bovine ephemeral fever.

The invention provides a method for obtaining a broadly neutralizing antibody (bNab), including screening memory B cell cultures from a donor PBMC sample for neutralization activity against a plurality of HIV-1 species, cloning a memory B cell that exhibits broad neutralization activity; and rescuing a monoclonal antibody from that memory B cell culture. The resultant monoclonal antibodies may be characterized by their ability to selectively bind epitopes from the Env proteins in native or monomeric form, as well as to inhibit infection of HIV-1 species from a plurality of clades. Compositions containing human monoclonal anti-HIV antibodies used for prophylaxis, diagnosis and treatment of HIV infection are provided. Methods for generating such antibodies by immunization using epitopes from conserved regions within the variable loops of gp120 are provided. Immunogens for generating anti-HIV1 bNAbs are also provided. Furthermore, methods for vaccination using suitable epitopes are provided.

The invention provides 3D tumor cell culture hydrogel as well as a preparation method and application thereof. The 3D tumor cell culture hydrogel is prepared from the following raw materials: polyvinylpyrrolidone and polyvinyl alcohol. The preparation method comprises the following steps: mixing a polyvinylpyrrolidone solution with a polyvinyl alcohol solution, adjusting the pH value, and freezing-unfreezing to obtain the 3D tumor cell culture hydrogel. The 3D tumor cell culture hydrogel provided by the invention does not need to be additionally added with a chemical cross-linking agent, and the material is high in biocompatibility and flexible in modulus adjustment, and can be well applied to a 3D in-vitro culture model of tumor cells.

The invention discloses a construction method and application of a cell senescence model, and belongs to the technical field of biology, firstly, cells are cultured in an embryonic-stage animal serum complete medium to a logarithmic phase for passage, then the cells are inoculated with an embryonic-stage animal serum complete medium, a newborn-stage animal serum complete medium or an adult-stage animal serum complete medium respectively, and the cell senescence model is obtained. The method comprises the following steps of: culturing and observing the cell state, and determining cell senescence by SA-beta-gal staining and detecting the transcriptional level of senescence-related genes when the cells grow slowly and become large and flat in comparison with an animal serum group in an embryonic period, so as to prove that the cell senescence model is successfully constructed. The cell senescence model is simple and convenient to operate, and the senescence induction process is simple, stable and rapid; the cell senescence model induced by newborn animal serum and adult animal serum simulates the senescence process of organisms of different age groups in vitro, and breaks through the limitation that cell senescence phenotypes are generated by oxidative stress treatment.

The present invention relates to the field of biotechnology, and particularly to a culture medium, a composition and a culture method for human NK cells. Through optimization, the present invention provides a culture medium, a composition and a culture method for NK cell culture. The culture medium, the composition and the culture method are used for NK cell culture, with clear culture components, simple culture steps, low cost, good amplification multiple of the cultured NK cells and high cell purity. At the same time, the cultured NK cells have good tumor cell killing ability and have good market promotion value.

The invention provides a culture method of avian encephalomyelitis virus, and belongs to the technical field of biology. According to the method, chicken lymphoma suspension cells are adopted to culture the avian encephalomyelitis virus, the titer of the obtained avian encephalomyelitis virus reaches 106.5-6.8 EID50 / 0.2 mL, and the method is short in culture period, high in production efficiency, low in cost, controllable in quality and capable of being used for large-scale industrial production.

The invention belongs to the technical field of cell mechanics biology experiment devices for health and rehabilitation engineering, and relates to a microfluidic device for endothelialization evaluation of a tissue engineering valve, which comprises a microfluidic chip, peripheral equipment, a signal acquisition and processing system and a feedback system. The biomechanical principle and the micro-fluidic chip technology are ingeniously combined, the control system, the mechanical loading device and the micro-fluidic chip are connected, a micro-fluidic endothelial cell culture cavity and a peripheral system are constructed, real-time accurate control over aortic valve blood flow is achieved through the PID control system, and the system is suitable for large-scale popularization and application. Shear stress applied to a tissue engineering valve endothelial cell culture area is generated, so that the real stress condition of the tissue engineering valve endothelial cells is simulated, and the method is used for analyzing and researching the influence rule of mechanical loading on the tissue engineering valve endothelial cells.

The utility model discloses a bioreactor bubble-free air interchanger which comprises a shell, an air inlet pipe extends into the shell, an air outlet port of the air inlet pipe extends into the shell, the top of the shell is connected with an exhaust pipe, an air inlet port of the exhaust pipe is located at the top of the shell, a liquid passing hole is formed in the cylinder wall of the shell, and the liquid passing hole is communicated with the air inlet port of the air inlet pipe. A filtering body for liquid to pass through is arranged on the periphery of the shell and is of a porous net structure. Compared with the prior art, the bubble-free air interchanger of the bioreactor has the advantages that the bubble-free air interchanger of the bioreactor is made of 316 stainless steel materials, is small in occupied space and easy to install and operate, can achieve bubble-free air interchange in the bioreactor, eliminates bubble damage to cells while meeting the requirement of a culture system for oxygen concentration, and improves the air interchange efficiency of the bioreactor. The device is suitable for the culture process of mammalian cells sensitive to shear force. The device is flexible in application, can be adapted to bioreactors of different specifications, and is easy for process development and amplification.

The invention discloses a virus-like plasmid and application thereof. The virus-like plasmid is characterized in that mIL-21 and 4-1BBL coding genes are inserted into a lentivirus plasmid; the amino acid sequences of the coding genes of the mIL-21 and the 4-1BBL are respectively as shown in SEQ ID NO. 1 and SEQ ID NO. 2. Virus-like particles (VLPs) prepared from the virus-like plasmids are added into an NK cell culture solution, and the VLPs can synergistically stimulate the NK cells, promote the rapid growth of the NK cells (the amplification multiple reaches 1850 times after 18 days of culture), and can improve the purity (about 92%) and the killing rate (when the efficiency-target ratio is 2: 1, up to 80%) of the NK cells.

The invention discloses a disinfection and sterilization device of a cell incubator and a use method of the disinfection and sterilization device, and belongs to the technical field of disinfection and sterilization of cell incubators. An electric push rod is controlled to jack up a movable seat to slide in a corresponding rectangular sliding groove, and a supporting frame is jacked out of a storage groove, so that the supporting frame rotates between the corresponding movable seat and a fixed seat; jacking the disinfection light plate to rotate at the bottom of the turntable until the disinfection light plate rotates at a proper angle at the bottom of the turntable, stopping the electric push rod from continuously pushing the movable seat, fixing the disinfection light plate at the bottom of the turntable at a specific angle, closing the magnetic suction box door, controlling the rotating motor to rotate, and opening the disinfection light plate at the same time; under the meshing transmission action of a bevel gear I and a bevel gear II, a rotating disc is controlled to drive a disinfection light plate to rotate in the cell culture box body, so that disinfection light rays emitted by the disinfection light plate irradiate each corner in the cell culture box body, and the interior of the cell culture box body is disinfected and sterilized.

The invention discloses a sleep-promoting nasal spray based on mesenchymal stem cell-derived exosomes and preparation and application thereof.The preparation method comprises the steps that 1, mesenchymal stem cells separated from umbilical cord tissue are cultured to P3-P5 generations, cell culture supernate is collected, cells are digested, and then cell debris and macromolecular impure protein are removed through pretreatment; (2) filtering and concentrating the pretreated cell culture supernatant and replacing a part of solution; (3) carrying out degerming filtration on the solution obtained in the step (2) through a 0.22 [mu] m filter membrane to obtain an MSC-Exos solution; and (4) adding a stabilizer with a proper concentration into the MSC-Exos solution to obtain a compound MSC-Exos solution, namely the sleep-promoting nasal spray. The product is a cell-derived natural active substance, can significantly improve sleep, and does not involve any pharmaceutical ingredient.

An anti-BDCA-2 antibody, a preparation method therefor, and a use thereof, relating to the field of antibodies. The preparation method for the anti-BDCA-2 antibody comprises: carrying out ELISA detection and screening on a hybridoma cell culture supernatant and an antigen protein to obtain a positive clone, and subcloning the positive clone to obtain the anti-BDCA-2 antibody. The anti-BDCA-2 antibody comprises a heavy chain and a light chain, wherein a variable region of the heavy chain and a variable region of the light chain are selected from amino acid sequences as shown in SEQ ID NOs: 1-22. The anti-BDCA-2 antibody and related drugs can be used for inhibiting the production of IFN-α by pDCs and preventing the over-activation of the pDCs. The anti-BDCA-2 antibody can effectively inhibit the activation of pDC cells and the production of IFN-α, and has great potential in resisting virus infection and preventing certain autoimmune diseases.

The invention discloses a preparation method of a cochlear organ chip integrated with a blood labyrinth barrier, and belongs to the field of biomedical engineering. The preparation method of the simulated blood labyrinth barrier comprises the following steps: constructing an endothelial cell culture cavity and a pericyte culture cavity, and separating the endothelial cell culture cavity and the pericyte culture cavity through a simulated basement membrane; and respectively inoculating percutaneous cells and endothelial cells into a percutaneous cell culture cavity and an endothelial cell culture cavity, and culturing to obtain the simulated blood lost barrier. The chip can effectively simulate the structure and the function of the BLB, and the barrier integrity of the BLB is evaluated through TEER (trans-epithelial electrical resistance) measurement and an apparent permeability coefficient. The invention also relates to application of the chip in NIHL drug screening, in particular to drug evaluation for oxidative stress induced inner ear organ injury. Through the platform, the protection effect of the candidate drugs on the TBHP-induced oxidative stress injury of the inner ear organs can be evaluated, and a new tool and thought are provided for developing novel NIHL treatment drugs.

The invention belongs to the technical field of microfluidic organ chips, and particularly relates to an anti-cancer drug screening chip and method based on cancer cell migration and angiogenesis, the organ chip comprises at least one monitoring culture module, and the monitoring culture module comprises a vascular endothelial cell culture layer, a PET porous membrane layer and a tumor cell culture layer which are fixedly connected in sequence; the vascular endothelial cell culture layer comprises a vascular endothelial cell culture chamber, and two sides of the vascular endothelial cell culture chamber are respectively communicated with a liquid inlet I and a liquid outlet I; the tumor cell culture layer comprises a tumor cell culture chamber, one side of the tumor cell culture chamber is communicated with a liquid inlet II, the tumor cell culture chamber and the vascular endothelial cell culture chamber are correspondingly arranged and communicated, and the PET porous membrane layer is arranged between the vascular endothelial cell culture chamber and the tumor cell culture chamber; one side of the migration chamber is communicated with the other side of the tumor cell culture chamber, and the other side of the migration chamber is communicated with a liquid outlet II.

The invention discloses a bioreactor multi-parameter adaptive control system and method based on machine learning, and belongs to the technical field of bioreactor intelligent control, and the bioreactor multi-parameter adaptive control system comprises a bioreactor hepatocyte culture unit, a data acquisition and control unit and a machine learning intelligent decision-making unit. And the data acquisition and control lower computer controls operation variables such as gas supply, sampling and perfusion in the culture unit execution mechanism, collects culture process data returned by the parameter detection module, constructs a data-driven cell state prediction model by utilizing machine learning, and optimizes model parameters and operation variables based on a prediction result to realize closed-loop control. The method can overcome the defect that there is no precise mechanism model for hepatic cell culture, and multi-parameter collaborative optimization can be realized only depending on process data, so that the state of the hepatic cells is sensed in real time, full-cycle optimization of cell viability and functions is realized, and the activity and density of hepatic cell culture are effectively improved.

The disclosure relates to growing cells, directing cells to grow into specified cell types, genetically and physically manipulating cells, and addressing one or more individual cells within a mixed cell population. Aspects of the disclosure relate to vectors useful to induce developmental changes in cells, in which those vectors have a temporal component. Vectors of the disclosure encode a controllable, temporal series of events. Once the vectors are delivered into target cells, a series of discrete and different genetic events may be induced. The disclosed methods generally provide for the temporal encoding of multiplex genetic effectors in vector format for cell state transitions.

A hybridoma cell strain 28F6 capable of secreting a high-titer anti-PRRSV monoclonal antibody is prepared and is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number of the hybridoma cell strain 28F6 is CCTCC NO: C2022373. The antibody titer in the cell culture supernatant of the hybridoma cell strain reaches 1: 640, the hybridoma ascites titer can reach 1: 4.096 * 10 < 5 > which is far higher than that of other prepared hybridoma cell strains, the monoclonal antibody secreted by the hybridoma cell strain 28F6, the PRRSV strain and the PRRSV vaccine attenuated strain obtained through passage have good reaction activity, and the monoclonal antibody and the PRRSV strain secreted by the hybridoma cell strain 28F6 have good immunogenicity. The probe has broad-spectrum reaction activity on the American type PRRSV and can be used for detecting the American type PRRSV.

The present invention relates to a method for rapidly preparing chimeric antigen receptor T cells (CAR-T), comprising the steps of: 1) contacting T cells or a cell population comprising the T cells with a stimulating agent to activate the T cells to obtain activated T cells or a cell population comprising the activated T cells; 2) contacting the activated T cell or a cell population comprising activated T cells with a nucleic acid molecule encoding a chimeric antigen receptor (CAR), thereby obtaining a CAR-T cell expressing the CAR; and (3) harvesting the CAR-T cells, culturing for 18-24 hours, harvesting after the culture is finished, and freezing or not freezing. According to the method, the total preparation time of the CAR-T cells is greatly shortened, and meanwhile, the amplification and tumor killing capabilities of the CAR-T cells are remarkably improved.

The invention discloses a culture method for improving proliferation and differentiation capacity of NK (Natural Killer) cells based on angelica sinensis-astragalus membranaceus exosomes, which comprises the following steps: firstly extracting exosomes in angelica sinensis and astragalus membranaceus, then adding the exosomes into an NK cell culture system according to a specific concentration, and regulating and controlling an NK cell signal channel by virtue of active ingredients carried by the exosomes so as to improve the proliferation and differentiation capacity of the NK cells. Therefore, the proliferation rate and differentiation maturity of the NK cells are remarkably improved. Experimental results show that compared with a traditional culture method, the proliferation multiple of the NK cells cultured through the method is increased by 28.99%-33.69%, the proportion of differentiated mature cells is increased by 15.91%-28.30%, and the cell killing activity is not remarkably reduced. The method can be widely applied to the field of immune cell treatment, provides a high-quality NK cell source for clinic, effectively solves the problems of slow cell proliferation rate, low differentiation maturity, easy activity reduction after large-scale culture and the like in traditional NK cell in-vitro culture, and has important clinical application value and industrialization prospect.

The invention relates to the technical field of vaccines, in particular to an activity detection method of an HPV (human papilloma virus) trivalent therapeutic vaccine. The method comprises the following steps: inoculating and infecting BEAS-2B cells by adopting constructed HPV16 / 18 / 52 type E6 / E7 antigen-loaded non-replicating type 5 adenovirus, culturing, extracting total protein, sequentially performing SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis), western blot detection, immunoreaction and developing imaging, and detecting the fusion protein; the transfection complex number MOI of the constructed adenovirus is 15-25, and the convergence degree of BEAS-2B cells is greater than 80%; the membrane transfer current of the western blot method is 220-280 mA, and the time is 50-80 minutes. The method avoids the problem of inaccurate detection results caused by obvious difference of biological activities in different animal species, has good accuracy, stability and durability, is simple in steps and low in cost, and can be effectively popularized and used.