85results about How to "Extend incubation time" patented technology

Multi-directional microfluidic devices and methods for using the same are provided. Aspects of the present disclosure include microfluidic devices that include a chamber having a separation medium, a first binding medium, and a second binding medium. In addition, the devices include a flow field element configured to subject the chamber to two or more directionally distinct flow fields. Methods of using the devices, as well as systems and kits that include the devices are also provided. The devices, systems and methods find use in a variety of different applications, including diagnostic, research and validation assays.

The invention discloses a method for separating and culturing primary chicken hepatocytes. According to the method, EGTA (Ethylene Glycol Tetraacetic Acid) with low hepatotoxicity is added into a perfusate A to loosen the junction between hepatocytes, so that the dispersion degree of the to-be-separated hepatocytes is increased; double resistance is added into the perfusate A, a perfusate B and D-Hanks flushing fluid, so that the pollution is further prevented; and circulating perfusion digestion is adopted during digestion perfusion, so that the perfusion cost is greatly reduced. A serum-free cultural method of the chicken hepatocytes is established; compared with a traditional serum cultural method, the method disclosed by the invention has the same effect on the aspects of cell quantity, activity, function and the like, so that the problems caused by the existence of serum are solved and a foundation is laid for the substance metabolism study and biological product preparation on the basis of the chicken hepatocyte culture.

The invention discloses an application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1), namely promoting proliferation and delaying senility of mesenchymal stem cells. The experimental result shows that the SIRT1 is needed by the in vitro proliferation of the mescenchymal stem cells, and the condition is verified by MSC (mesenchymal stem cells from marrow) and MSC from fat, and the proliferation and the senility of the MSC can be regulated by regulating the expression level of SIRT1 in the MSC. The invention has very important significances on the determination of the regulation mechanism of the proliferation and the senility of the MSC, the prolonging of the service life of the MSC, and the histocyte substitution and repairing treatment based on the MSC, and is broad in application prospect.

The invention relates to a novel dual-layer composite bone tissue engineering scaffold and a preparation method thereof. The dual-layer bone tissue engineering scaffold comprises an outer-layer bacterial cellulose membrane and an inner-layer PLGA / MWNTs electrostatic spinning porous scaffold. The synthetic material PLGA, a natural ingredient bacterial cellulose and an inorganic ingredient multi-wall carbon nanotubes are compounded together to form the composite material by using a vacuum freeze drying process and a high-pressure electrostatic spinning method. The materials have complementary advantages, and an ideal dual-layer composite bone tissue engineering scaffold of PLGA / MWNTs / bacterial cellulose, which is excellent in performance and relatively cheap, is built. The dual-layer composite bone tissue engineering scaffold of the PLGA / MWNTs / bacterial cellulose provided by the invention is simple in preparation method, and mild in condition, has good cell biocompatibility, accords with the requirements of application inside a living body, and has a good application prospect as a novel bone tissue engineering scaffold.

The invention relates to a culture medium of an induced haplobiont for culturing eggplant anther and a method of the culture medium, and belongs to the technical field of plant biology. The method for cultivating the eggplant anther provided by the invention comprises the following steps: with the eggplant anther as a material, carrying out anther culture to induce callus; carrying out differentiation culture on the callus; carrying out rooting culture on differentiated seedlings; carrying out acclimatization and transplanting; and carrying out haplobiont doubling germination, so as to finish regeneration of an eggplant plant. The culture medium provided by the invention is low in cost and good in culture effect; the method for culturing the eggplant anther by virtue of the culture medium provided by the invention has the advantages of high callus induction rate, simple culture method and short culture period; the culture medium and the eggplant anther culture technology provided by the invention are applied to eggplant genetic breeding; the excellent genotype individual bodies of the eggplant are greatly enriched; the breeding period is obviously shortened; and a reliable technical support is provided for variety improvement of the eggplant.

The invention provides an application of a compound microorganism bacterium agent in treatment of petrochemical sewage and sludge. The application comprises the following steps: (1) preparing a biological carrier, to be specific, adding glutaraldehyde to polyphenylene oxide, suspending, stirring, washing, then adding a PBS (Phosphate Buffer Solution) containing CaCl2 and standing; (2) carrying out enlarging cultivation of the compound microorganism bacterium agent; (3) putting the compound microorganism bacterium agent subjected to enlarging cultivation, obtained in the step (2), in the biological carrier prepared in the step (1); and (4) adding the biological carrier loaded with the compound microorganism bacterium agent, obtained in the step (3), to the petrochemical sewage sludge, wherein the compound microorganism bacterium agent continuously increases to a stable period and forms a biological membrane on the surface of the biological carrier, and the biological membrane metabolizes, adsorbs, absorbs, digests and resolves organic pollutants and heavy metals in petrochemical sewage and sludge, so that the petrochemical sewage and sludge are converted into stable harmless materials. Through research, the applicant finds that the compound microorganism bacterium agent can be used for treating the petrochemical sewage and sludge very well.

The invention discloses a method for reprogramming umbilical cord mesenchymal stem cells into liver cells. The method comprises the following steps of obtaining umbilical cord mesenchymal stem cells;carrying out pretreatment on the umbilical cord mesenchymal stem cells for two days; carrying out liver formation induction on the pretreated umbilical cord mesenchymal stem cells for seven days by using a liver formation induction composition; carrying out primary maturation induction on the umbilical cord mesenchymal stem cells subjected to liver formation induction treatment by using a first maturation induction composition for seven days to obtain liver cells; and carrying out second maturation induction on the liver cells for 12-20 days by using a second maturation induction composition to obtain the functional liver cells. Liver organoids comprise acellular matrix hydrogel, stem cells and liver cells, wherein the stem cells and liver cells being distributed in the acellular matrix hydrogel. According to the reprogramming method, the step of secondary maturation induction is added, so tat the culture time is prolonged, and the induction efficiency is remarkably improved. The liverorganoids provided by the invention have ultra-low immunogenicity and the possibility of cross-species transplantation.

The invention discloses application of an artemisia verlotorum extract to prevention and control of panax pseudoginseng black spot, belonging to the technical fields of plant pest prevention and control. A preparation method of the artemisia verlotorum extract comprises the following steps of (1) naturally airing after cleaning fresh stems and leaves of disease-free artemisia verlotorum, sieving after smashing the fresh stems and leaves of the disease-free artemisia verlotorum, then, mixing the stem powder and the leaf powder, and placing the mixed powder in a dry vessel; (2) adding ethanol into the artemisia verlotorum powder obtained in the step (1) to filter for the first time to obtain a filtrate, adding ethanol into the obtained filter residue again to filter for the second time, repeatedly filtering twice, and then, mixing the filtrates obtained in three times; and (3) carrying out reduced-pressure condensation and ultraviolet sterilization on the filtrate obtained in the step (2), and then, keeping the constant volume by using sterile water to obtain the artemisia verlotorum extract. The artemisia verlotorum extract disclosed by the invention has a favorable antibacterial effect for pathogenic bacteria of panax pseudoginseng black spot.

The invention relates to the field of microalgae symbiotic bacteria isolation, specifically speaking, relates to a microalgae symbiotic bacteria isolation medium and separation method, and a high-throughput screening method for key bacteria affecting the growth of microalgae. Improvement can be performed based on a common marine bacterial isolation medium - Zobell 2216E marine broth bacterial culture medium, so that the culture medium for increasing the culturable bacterial species of microalgae phycosphere symbiotic bacteria can be provided, and abundant strain resources can be provided for the screening of key bacteria affecting the growth of microalgae. Through the high-throughput rapid screening method for significantly promoting or inhibiting algae growth bacteria, bacteria having keyinfluences on the growth of the microalgae can be discovered in a short time.

The invention discloses a liquid rapid-propagation method for rhizoma bletillae seeds. The method comprises the following steps: (1) explant selection and disinfection; (2) seed germination and young-seedling culture; (3) adult-seedling culture; and (4) seedling hardening and transplantation. The step of seed germination and young-seedling culture concretely comprises the sub-step of culturing the rhizoma bletillae seeds in a liquid medium, wherein culture conditions are as follows: temperature is 26+ / -2 DEG C; illumination intensity is 3000 to 3500 Lx; illumination time is 14 to 16 hours / day; and culture time is 45 to 60 days. The liquid rapid-propagation method for the rhizoma bletillae seeds, provided by the invention, can realize industrialization of rhizoma bletillae seedling propagation and obtains rhizoma bletillae seedlings meeting the needs of large-scale production in a short time; meanwhile, the method also has the characteristics of low cost, neat seedling growing, high seedling-hardening domestication survival rate, etc.

A farming windrow machine and method for sowing a second crop before the first crop is harvested. The windrow machine is preferably hitched to a tractor and is followed by an agricultural machine to sow a field immediately after the crop nearly ready for harvested is cut. The machine omprises a frame supporting means for aligning crop upstanding in the field and mower means for cutting the aligned crop. The machine further comprises a conveyor belt mounted on a pair of spaced rollers, the conveyor belt cantilevered over a side of the windrow machine for transporting and piling the cut crop over and away from one side of the machine, such that a sower machine may follow the windrow machine to sow a second crop in the wake of the first cut crop, thereby increasing crop cultivation days and improving farm turnover.

The purpose is to provide the truffle mycelium and fruiting body cultivated by the medium that are developed for cultivating the truffles with a very high radical oxygen scavenging capacity. A medium for cultivating truffles includes a mixture of at least one carbohydrate source selected from a group consisting of rice bran, wheat bran, and glucose, and at least one edible ingredient selected from a group consisting of burdock, nuts, and berries. A weight ratio of the edible ingredient to the carbohydrate source is between 0.5 and 5. As for one of the ingredients, it is preferable that the burdock is granulated. The medium may exist in a liquid state. By injecting truffle seeds for cultivation into such a medium, the truffle mycelium and fruiting body with the high reactive oxygen scavenging capacity could be provided.

The invention discloses a new strain of pseudoxanthomonas beigongshangensis and an application of the new strain. The strain is pseudoxanthomonas beigongshangensis, and the preservation number of thepseudoxanthomonas beigongshangensis in the China General Microbiological Culture Collection Center is CGMCC NO.19208. The tests prove that the invention provides a new strain of pseudoxanthomonas beigongshangensis, and the strain is separated from pit mud of white wine, can generate aromatic flavor substances, and can be used for fermenting and brewing wine.

The invention belongs to the field of biotechnology, and discloses an in-vitro cell culture method for primary B cell acute lymphoblastic leukemia. According to the in-vitro cell culture method, primary B-ALL cells and OP9 cells are co-cultured so as to maintain the survival rate and proliferation of primary B-ALL cells and greatly relieve the differentiation and senescence of the B-ALL cells. The primary B-ALL cells cultured by adopting the culture method can be proliferated, the cell status is good, and the heterogeneity of tumor cells can be fully embodied. The method is simple, convenient, and economic to operate, and has good application prospects.

The invention provides an application of a compound microorganism bacterium agent in treatment of petrochemical sewage and sludge. The application comprises the following steps: (1) preparing a biological carrier, to be specific, adding glutaraldehyde to polyphenylene oxide, suspending, stirring, washing, then adding a PBS (Phosphate Buffer Solution) containing CaCl2 and standing; (2) carrying out enlarging cultivation of the compound microorganism bacterium agent; (3) putting the compound microorganism bacterium agent subjected to enlarging cultivation, obtained in the step (2), in the biological carrier prepared in the step (1); and (4) adding the biological carrier loaded with the compound microorganism bacterium agent, obtained in the step (3), to the petrochemical sewage sludge, wherein the compound microorganism bacterium agent continuously increases to a stable period and forms a biological membrane on the surface of the biological carrier, and the biological membrane metabolizes, adsorbs, absorbs, digests and resolves organic pollutants and heavy metals in petrochemical sewage and sludge, so that the petrochemical sewage and sludge are converted into stable harmless materials. Through research, the applicant finds that the compound microorganism bacterium agent can be used for treating the petrochemical sewage and sludge very well.

The invention relates to a culture medium for ricefield eel germline stem cells. The culture medium comprises a basic culture medium and FBS, wherein the concentration of the FBS is 5-8%. The invention also relates to a method for separating and culturing the ricefield eel germline stem cells. The method comprises the step of culturing the ricefield eel germline stem cells by using the culture medium. According to the culture medium disclosed by the invention, the adherence efficiency of the ricefield eel germline stem cells can be improved, differentiation of the ricefield eel germline stem cells is prevented, and the proliferation function of the ricefield eel germline stem cells is not influenced. By the method disclosed by the invention, ricefield eel female germline stem cells and male germline stem cells which have higher purity can be separated and obtained in five days by using gonad tissues of ricefield eels. The obtained germline stem cells have typical stem cell morphology,typical stem cell clone can be formed, alkaline phosphatase staining is positive, and related molecular markers of the stem cells are also highly expressed. After the germline stem cells obtained by the method disclosed by the invention are cryopreserved, the germline stem cells can be successfully recovered.

The invention provides an isolation and culture method for primary rat colonic smooth muscle cells. With the isolation and culture method provided by the invention, the acquired cells are great in gross quantity and high in both isolation degree and motility rate; the probability of contamination by bacteria, fungi and other cells is low; and the method can realize subculturing, so a good cell culture scheme is provided for related research based on culture of rat colonic smooth muscle cells.

The invention discloses a hybridoma cell serum-free medium, which comprises the following raw materials: alanine, arginine, aspartic acid, asparagine, cystine, cysteine, glutamine, glutamic acid Amino acid, glycine, histidine, etc., calcium chloride, magnesium sulfate, potassium chloride, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium bicarbonate, sodium hydroxide, ferrous sulfate heptahydrate, magnesium chloride hexahydrate, sulfuric acid Copper, sodium pyruvate, manganese dichloride, sodium selenite, ammonium molybdate, etc., nicotinamide, p-aminobenzoic acid, folic acid, ribose, uracil, lipoic acid, linoleic acid L, cholesterol, dexamethasone, Estradiol progesterone, testosterone, etc. The medium of the present invention does not contain serum components at all; the purity of the produced antibody is greatly improved; the medium has high amplification speed and high antibody expression ability; the medium can maintain cell activity for a long time, and the hybridoma cell culture time is greatly improved Extending can greatly extend the production time and increase the antibody production.

The invention relates to a preparation method and application of a low-grade attapulgite modified material. The preparation method of the low-grade attapulgite modified material is characterized in that low-grade attapulgite powder and an ethylenediamine aqueous solution with the concentration of 1%-15% (v / v) are mixed according to a solid-liquid ratio of 50-200 g / l, then are subjected to hydrothermal reaction at 180-220 DEG C for 4-24 hours, then are cooled to room temperature, and solid precipitates are separated and dried to obtain the low-grade attapulgite modified material. The low-gradeattapulgite modified material has strong adsorption effect on cadmium and other heavy metal ions and / or forms insoluble mineral salts, so that the low-grade attapulgite modified material changes intoa form unavailable to crops, thereby blocking the transformation of the cadmium and other heavy metal ions into the crops and reducing the biotoxicity thereof under the condition that the cadmium andother heavy metal ions are not removed from soil; moreover, the clay mineral is proved to have better water and fertilizer conservation characteristics, and finally the purposes of pollution control and ecological restoration can be achieved.

The invention discloses a culture medium for culturing astroglia cells in vitro and a culture method and belongs to the field of cell culture. According to the culture medium, an epidermal growth factor, a fibroblast growth factor, SAG and Purmorphamine are added into an astroglia cell culture medium, so that the in-vitro culture time of the astroglia cells can be remarkably prolonged, the numberof time of passage is increased, the purity is improved and original properties of the astroglia cells can be kept. The inventor researches and finds out that the culture medium disclosed by the invention can be used across generations, and the astroglia cells are passed to 1 to 3 generations. The purity of the astroglia cells, which are cultured to 12 generations, is greater than 95 percent and the original properties of the astroglia cells can be kept.

The invention relates to the use of an agent such as an antibody capable of reacting with PrP in the prevention of prion replication in a subject, in the treatment or prevention of prion infection, in the treatment or prevention of neuropathology associated with prion infection or in the preparation of a medicament for the treatment or prevention of prion disease. Furthermore, the invention relates to methods of treatment of prion disease, methods of inhibiting prion replication and antibodies for use in such methods.

The invention discloses a method for preparing Saccharomyces cerevisiae cell culture secretion and the function of the secretion in DNA virus resistance and acceleration of hepatocyte growth, pertaining to the field of bio-pharmaceutics. The following method is adopted to prepare Saccharomyces cerevisiae cell culture secretion: monoclonal colony of Saccharomyces cerevisiae is selected and inoculated into 5ml PD culture medium, pendulated under 30 DEG C for a night; 1 ml is inoculated into 500ml PD culture medium, and pendulated under 28-30 DEG C for 40-60h; 0.2-micron filter membrane is used for filtration sterilization to achieve the product. The invention has the advantages that Saccharomyces cerevisiae cell culture secretion after cultivation by culture medium and purification has eminent function in DNA virus resistance and acceleration of hepatocyte growth, and is a novel medicine to cure hepatitis B and liver-injury diseases.

The invention discloses a device and a method for in-vitro preservation and culturing of a blood vessel, and belongs to the field of biomedical engineering. The device for the preservation and culturing of the blood vessel consists of a blood vessel culture cavity, a circulating system pressure adjuster, a blood vessel deformation adjuster, a pump, a damping adjuster, a liquid storage chamber, anda computer control system, wherein the blood vessel culture cavity, the circulating system pressure adjuster, the pump, the damping adjuster and the liquid storage chamber are connected by pipelinesto form a fluid circulating system; the blood vessel deformation adjuster is used for controlling the deformation of the blood vessel in the blood vessel culture cavity; the circulating system pressure adjuster, the blood vessel deformation adjuster and the pump are controlled by the computer control system; expect the computer control system, the whole device can be placed into a CO2 (carbon dioxide) culture box to perform blood vessel preservation and culture. The device has the advantage that the function of the physical environment on the blood vessel in vivo can be simulated, and the functions of flow rate, pressure, deformation and temperature are applied to the blood vessel or the simulated blood vessel, so that the blood vessel can be dynamically preserved and stored in vitro for alonger time under the bionic simulating state.

The invention provides an escherichia coli for high expression of a foreign protein. A genotype of the escherichia coli lacks a prc gene and carries a mutated spr gene and a mutated ilvG gene; the mutated spr gene is as follows: the 174th position encoded by a spr gene is mutated to arginine or lysine by tryptophan; and the mutated ilvG gene is as follows: two bases of 'TA' are inserted at the 981bp position of the ilvG gene. The escherichia coli achieves a stable period of growth of a escherichia coli strain in the high-density fermentation process, and the degradation of the foreign proteinsecreted and expressed in a periplasmic space of the escherichia coli is inhibited, the high expression of the foreign protein is achieved, and the expression level is improved.

The invention belongs to the technical field of condiments, and particularly relates to a preparation method of calcareous black bean soy sauce. The calcareous black bean soy sauce is prepared from main raw materials as follows: black beans, pig elbow bone, wheat and coarse bran, wherein a ratio of the black beans, the pig elbow bone, the wheat and the coarse bran in parts by weight is (4-12):(2-6):(1-3):(1-4.5), and preferably, the ratio is 4:2:1:1.5. The preparation method of the calcareous black bean soy sauce comprises steps as follows: raw material processing; starter-making based on circulating ventilation; first solid fermentation and later liquid fermentation; oil sprinkling and sterilization; packaging, testing and storage; packaging and sampling test. Technological parameters are strictly controlled in the whole soy sauce preparation process, and are different from those just set in a certain large range for soy sauce on sale in the market. The calcareous black bean soy sauce prepared with the method is delicious in taste and has the function of calcium supplementation.

The invention discloses stem cell culture equipment. The stem cell culture equipment comprises a culture box; the front surface of the culture box is provided with a sealing cover; two clamping grooves are formed in the front surface of the culture box; the inner side of the sealing cover is fixedly connected with two fasteners; an installation box is fixedly installed in the culture box; the inner bottom part of the installation box is fixedly provided with a motor; the inner bottom part of the installation box is fixedly provided with a fixed frame body; an output end of the motor penetratesthrough one side of the fixed frame body and is fixedly connected with a first bevel gear; and the outer surface of the first bevel gear is in engaged connection with a second bevel gear. A culture dish is placed in a fixed holding groove; the culture dish can rotate along when a second rotating disc rotates, so that a culture solution in the culture dish is in a circular motion state, thus preventing the culture solution from being solidified when being statically placed, so that stem cells are kept active for a long time, and then the culture time is prolonged.

A Petri dish includes a receptacle and a matching lid, which both have a shape of revolution and are each delimited by a bottom wall and at least one peripheral rim. This also includes fingers and complementary locking elements. The receptacle and the lid, when rotated relative to each other, are able to occupy three or four different positions, including at least a “first position” and an “intermediate position”. This dish is distinguished by the fact that the locking elements carry at least one blocking member which blocks the fingers in the “intermediate position” and which opposes the return to the “first position”.

The invention relates to a CAR-T cell, a preparation method thereof and a medicine. Negative T cells which fail to transduce CAR genes are removed through culture and purification operation of specific steps, T cells with puromycin resistance genes are reserved, and the proportion of CAR positive T cells is greatly increased. More importantly, experimental comparison shows that although the culture time of the CAR-T cells obtained through culture and purification operation of the specific steps is prolonged, the side effects of the CAR T cells in application are remarkably reduced, for example, the secretion level of inflammatory cytokines IL-6 and other proinflammatory related cytokines is reduced; Th17 cell subgroup components are reduced, and IL-17 / IL-17A secretion is reduced; anti-inflammatory related cytokine secretion is increased; and the killing capacity on tumor cells in vitro and in mice is greatly enhanced, and the survival time of the mice is prolonged.

The invention discloses a method for promoting cells to differentiate into female genital cells based on overexpression of Figla gene. The Figla gene has the nucleotide sequence shown in SEQ ID. NO.1. The Figla gene is labeled and is used for transfecting pluripotent stem cells so as to promote the pluripotent stem cells to differentiate or transdifferentiate into the female genital cells. After pDsRed1-N1-Figla transfercts mESCs (Mouse Embryonic Stem Cells), RFP (Red Fluorescent Protein) is expressed in noble cells at the cell cloning margin and not expressed in the smooth position at the cloning margin; and the transfected mESCs grow in a flake shape after passage, and most of the transfected mESCs do not form cloning, indicating the Figla gene promotes the differentiation of mESCs.

The invention discloses a preparation method of liver organoid. The preparation method comprises the following steps: uniformly mixing acellular matrix hydrogel, stem cells and liver cells by using a serum-free culture medium; conducting stationary culture in a culture container until the acellular matrix hydrogel is solidified; and after it is determined that cells in the solidified acellular matrix hydrogel survive, continuing culture to obtain the liver organoid. The liver organoid prepared according to the method of the invention has ultralow immunogenicity and has the potential in cross-species transplantation.