47 results about "Culture expansion" patented technology

The invention relates to methods of isolating and implanting mesenchymal stem cells for tissue repair or formation, without prior culture expansion of the mesenchymal stem cells. In particular, the invention relates to methods of isolating mesenchymal stem cells from bone marrow, for repairing or inducing formation of bone, without prior culture expansion of the mesenchymal stem cells. The invention further relates to an isolated, non-culurally expanded human adult mesenchymal stem cell population.

The invention discloses a complex bacterium preparation for fixing nitrogen, solubilizing phosphorus and potassium and rooting and a preparation method thereof. The preparation is mainly prepared by mixing the following raw materials in parts by weight: 5-20 parts of azotobacter vinelandii, 10-30 parts of bacillus megatherium, 10-30 parts of bacillus mucilaginosus, 5-15 parts of azotobacter chroococcum, 10-15 parts of glomus intraradices and 10-25 parts of glomus mosseae. The preparation method comprises the following steps: respectively carrying out high-density culture on each strain; grafting to a shake flask filled with a culture medium in a volume ratio of 15-30 percent for activated culture; grafting the cultured strains into a fermentation tank for fermentation culture expansion; dehydrating and drying the culture expanded single strains to prepare hypopus microorganism dried powder; and mixing to obtain the complex bacterium preparation for fixing nitrogen, solubilizing phosphorus and potassium and rooting. The complex bacterium preparation is used for industrial crops, fruit trees, food crops and the like, has the advantages of biological property, environmental friendliness, high yield and high quality in comparison with existing products, and cannot pollute soil.

The invention discloses a fermentation process of lucid ganoderma mycelium. The fermentation process includes a lucid ganoderma mycelium seed culture stage and a culture expansion state, wherein the culture expansion stage continues for 5-7 days, and lighting processing is conducted on the lucid ganoderma mycelium every day by cooperating with a variable-temperature environment and specifically comprises the following steps that culture is performed under the lighting condition for 4-16 hours, and culture is performed under the variable-temperature condition for 1-3 hours, wherein the variable-temperature condition is light shielding and high or low temperature keeping. Due to the fact that lighting and variable temperature are combined for processing, high-level accumulation of the intracellular triterpene content of lucid ganoderma is achieved, cheap, safe and easy operation can be achieved on the premise of not adding any harmless allogenic material, the intracellular ganoderma triterpene content can be remarkably increased, and the fermentation process is applicable to industrial production of ganoderma triterpenes.

The invention relates to the technical field of applied biology, and aims to provide a bacillus subtilis strain and application of the bacillus subtilis strain in the preparation of a deodorizing microorganism. The bacillus subtilis strain is stored in China General Microbiological Culture Collection Center (CGMCC) of China Microbiological Culture Conservation Committee, and the conservation number is CGMCCNo.13714. The adopted technical scheme includes the steps of inoculating the bacillus subtilis CGMCC.13714 to an activated culture medium to obtain an activated strain, and inoculating the activated strain to an expanded fermentation medium for culture expansion to obtain a bacterial solution; inoculating the bacterial solution to faeces, and keeping the temperature at 20-35 DEG C. The novel Bacillus subtilis strain can be applied to the biological deodorization of faeces, and the deodorization effect of single use is stable and slightly influenced by the temperature; effective deodorization can be conducted by an in-situ deodorizing method, the operation is simple and easy, no supporting equipment and technology are needed, and the investment scale is small.

The invention relates to a method for reducing egg cholesterol by utilizing lactobacillus paracasei KL1 living bacterium agent of bile salt producing hydrolase. The method is suitable for production low-cholesterol eggs in poultry and animal husbandry. The method utilizes a lactobacillus paracasei KL1 strain of the bile salt producing hydrolase screened from Tibetan kefir to prepare the living bacterium agent through activation, culture expansion, high-density fermentation, centrifugal concentration, freeze-drying protective additive adding, pre-freezing, freeze drying and other processes; then the living bacterium agent with the living bacterium number of 1010 CFU / g or above is added to a basic ration for raising laying hens, the adding amount is the daily intake 106 CFU and 107 CFU of each hen, a feed containing the living bacterium agent is fed for 8-10 weeks, the cholesterol content in egg yolk can be reduced to be about 11%, and meanwhile the living bacterium agent has the effects of improving the laying rate of the laying hens, egg shell quality and relative egg yolk weight and reducing the fatty liver of the laying hens. The method is simple in operation, low in cost and suitable for large scale farming and production of low-cholesterol eggs.

The invention relates to a preparation method of a semi-solid micro-ecological preparation for aquaculture and belongs to the technical field of the aquaculture. The preparation method includes that byproduct bran, wheat middling and soy sauce residues of industrial and agricultural production are taken as a fermentation matrix, microbiological strains of bacillus natto CICC 10454, lactobacillus acidophilus CICC 21007, acetobacter aceti CICC 21684, saccharomyces cerevisiae CICC 1005 and candida utilis which have different characters and ecological activities are used for mixed fermentation toform a fermentation product high in viable count, protein and amino acid content, and the semi-solid micro-ecological preparation integrated with functions of water purification, pathogenic inhibition, intestinal repairing, liver protection, digestion promotion, disease prevention and growth promotion and the like is obtained via vacuum package. Characters of water and powder of the microbial preparation are combined with those of multi-strains effectively, so that the semi-solid micro-ecological preparation is long in storage life, high in activity and convenient to carry; the semi-solid micro-ecological preparation can be used directly by being sprayed or stirred with materials as well as used after culture expansion.

The invention provides a mycotoxin penicillic acid monoclonal antibody and a preparation method thereof. The preparation method comprises the following steps of coupling penicillic acid to carrier protein respectively so as to obtain complete antigen, immunizing a mouse with the complete antigen, adopting a hybridoma technique and obtaining the monoclonal antibody by cell fusion, screening, cloning, culture expansion and an animal in vivo induction ascites method. The obtained antibody has the titer of 1:2.05x10<5>, is of an IgG1 type, has no cross reaction rate with aflatoxin B1, zearalenone, T-2 toxin and Fumonisins, has the affinity constant of 1.54x10<8> L / mol, and can be used for developing ELISA kits for detecting the penicillic acid.

There is provided a method for the isolation and cultivation of mesenchymal stem / progenitor cells from umbilical cord blood, and also to a method for the differentiation of the umbilical cord blood-derived mesenchymal stem / progenitor cells into various mesenchymal tissues. The method include overlaying umbilical cord blood onto Ficoll-Hypaque solution; centrifuging the umbilical cord blood on the Ficoll-Hypaque solution to obtain a mononuclear cell layer; reacting cells obtained by monolayer culture of the mononuclear cells with antibodies to mesenchymal stem / progenitor cell-specific antigens for a predetermined period of incubation time; isolating only cells bound to their corresponding antibodies using a cell sorter; and cultivating the isolated cells, thereby obtaining mesenchymal stem / progenitor cells with high purity and excellent viability.

The invention relates to the biotechnology field and in particular to a CIK cell culture method. The method comprises the following steps: (1) heavy-suspending a mononuclear cell of peripheral blood in CIK culture solution, to obtain cell suspension; (2) adding the cell suspension into a culture bottle pre-enveloped by an anti-CD3 monoclonal antibody and retronectin, culturing in a culture box with 5% of CO2 saturation humidity in 37 DEG C of the temperature; (3) replenishing the CIK culture solution in the third day of the culturing; (4) in the sixth day of the culturing, after replenishing the CIK culture solution, executing the culture expansion; (5) replenishing the CIK culture solution in the ninth day of the culturing; and (6) culturing for 13-14 days, and harvesting the cells. The method is capable of remarkably improving the proliferation rate of the immune cell, especially the proliferation rate of CD3+CD56+ effector cell, and improving the killing activity thereof.

The invention discloses a fishery ocean microbial ecological preparation. The preparation comprises lactobacillus pantarum and pediococcus pentosaceus, wherein the microbial content ratio of lactobacillus pantarum to pediococcus pentosaceus is 2:1. The invention further discloses a preparation method of the fishery ocean microbial ecological preparation. The preparation method comprises a first step of activation and culture expansion of strains; a second step of preparation of a fermentation medium; a third step of fermentation cultivation; a fourth step of filtering and material discharging. A culture medium formula facilitating large-scale production is screened out for lactobacillus pantarum and pediococcus pentosaceus, the feed efficiency is greatly improved, metabolites of lactobacillus pantarum and pediococcus pentosaceus are synergistic, the efficiency of the metabolites of lactobacillus pantarum and pediococcus pentosaceus is much higher than that of a metabolite of lactobacillus pantarum or pediococcus pentosaceus in production application, and popularization and application of lactobacillus pantarum and pediococcus pentosaceus microbial ecological preparation for aquaculture are effectively promoted.

The invention provides an expansion culture method of CD8 T cells. The CD8 T cells are subjected to induced expansion culture in the presence of the following substances: K3EC cells and a peptide-fragment overlapping library of a target antigen; and the K3EC cells are K562 engineering cells expressing CD2 ligands CD58, NKG2D ligands MICA / B and CD137 ligands CD137L and are used for providing costimulatory receptor second signals activating the CD8 T cells, and the peptide-fragment overlapping library of the target antigen is used for providing T cell receptor (TCR) first signals activating theCD8 T cells. According to the method, quick and efficient expansion of the antigen (CMV-pp65) specific CD8 T cells; and the CD8 T cells obtained through expansion have high clinical application valuein prevention and treatment of CMV infection and treatment of CMV-related cancers.

The invention provides a cultivation method for and application of antrodiacamphorata mycelia. The cultivation method for antrodiacamphorata mycelia comprises the steps of: (1) culture expansion in shake flasks: inoculating antrodia camphorate strains to an inclined surface culture substrate for inclined surface culture, inoculating the antrodia camphorate strains to a liquid culture substrate after the inclined surface culture and performing primary shake flask culture, and then inoculating the antrodia camphorate strains to a liquid culture substrate for secondary shake flask culture; (2) culture expansion in seeding tanks: inoculating production shake flask second-level seeds into a fermentation culture substrate for culture expansion to obtain first-level antrodia camphorate strains; (3) culture expansion in fermentation tanks: inoculating the first-level antrodia camphorate strains into fermentation tanks with fermentation culture substrates for fermentation tank culture, and performing filtering to obtain antrodiacamphorata mycelia. Antrodiacamphorata mycelia produced by using the method have the advantages of short production period and high quality. Beverages prepared fromthe antrodiacamphorata mycelia have a good health care effect.

A method for improving the content of a characteristic aroma component ethyl propionate of Te-flavor Chinese spirits. Acid-proof propionibacterium with high yield of propionic acid is applied. After the amplification culture, propionibacterium liquid of volume 0.33%-1% is injected into a pit. The time for pit injection is controlled in the main fermentation period, namely the 15th day after pit opening of spirits sediment and the depth of pit injection is from 20 cm to 30 cm, the diameter of pit injection is 2.5 cm. Seed culture solution of propionibacterium is obtained through culture expansion in the invention. After the main fermentation, propionibacterium liquid is injected into the pit. Results show that the effect is obvious in the increase of content of ethyl propionate in spirit base generated in a pit with added propionibacterium liquid. The content of generated ethyl propionate is more than 2 times that of a blank pit. After tasting and evaluation, the increase in content of ethyl propionate creates a sweeter and softer taste and a more comfortable aroma of spirit base. Not only the quality of Te-flavor Chinese spirits can be improved, but also influences on yield are controlled in a reasonable scope after the pit injection time and the bacteria liquid amount are well controlled.

The invention relates to the technical field of plant fermentation beverages, and discloses a method for preparing a fruit enzyme through culture expanding fermentation with fruit juice as the culture medium.The method comprises the steps of cleaning and sorting, curing, basic fermentation, culture expansion, stable fermentation and filtration.The culture expanding fermentation method with fruit juice as the culture medium is free of additive and is purely natural.

The invention relates to a construction method of genetic engineering strains for improving quantity of soybean nodules. The construction method aims at solving the problems that when the conventional rhizobium japonicum genetic engineering strains are inoculated to soybean seedlings, the root nodule quantity of soybean plants and the yield increase of soybeans are small compared with those of the original starting strains. The construction method comprises the following steps of: performing activation and culture expansion on sinorhizobium fredii to extract genomic DNA (deoxyribonucleic acid); performing PCR (Polymerase Chain Reaction) amplification by virtue of a homologous sequence cloning method to obtain target gene dct A; performing sequencing identification on the target gene; constructing a prokaryotes expression vector pTR-Plac-dct A; and converting the prokaryotes expression vector into the sinorhizobium fredii to obtain the gene engineering strains. The quantity of the root nodules on the soybean plant with the transformed gene engineering strains is increased by 53.73%, and the yield of the soybeans is increased by 62.02%.

The invention discloses construction and application of an immortalized primary generation sheep lung cell line. Construction comprises the following steps that fresh primary generation sheep lung fibroblasts with the high viability are obtained through a Dispase-collagenase perfusion method; after being cultured for 24 h, under the condition that polybrene with the concentration being 5 ug / ml exists, the primary generation sheep lung fibroblasts are infected through concentrated and purified cell supernatant of recombinant lentivirus containing hTERT genes; after one week, puromycin with the concentration being 2 ug / ml is used for drug pressure screening, and under the condition that the polybrene with the concentration being 5 ug / ml exists, repeated infection is conducted one time; and after one week, the primary generation sheep lung fibroblasts are cultured through puromycin maintenance media with the concentration being 1 ug / ml till monoclonal cells appear and grow to be about 2.0 cm, the monoclonal cells are selected for culture expansion, and thus the immortalized sheep lung fibroblasts with a passage characteristic can be obtained.

The invention provides an environment-friendly synthesis method of a biological dust suppressant. The environment-friendly synthesis method of the biological dust suppressant comprises the steps of activation culture, culture expansion cultivation and culture fermentation cultivation; the principle is that working bacterial strains in a dormant state are awakened through the activation culture, working bacteria with absolutely superior number are obtained through the culture expansion cultivation, the working bacteria are enabled to take a fermented Landy culture medium as a medium through theculture fermentation cultivation, and surfactin with excellent surface activity and wettability is synthesized by secondary metabolism. According to the environment-friendly synthesis method of the biological dust suppressant, no extreme condition tolerance requirement is required for a synthesis device, the synthesis method is simple, the synthesized surfactin can significantly reduce the contact angle of a dust suppressant solution and dust at a very low concentration, the ability of wetting the dust is fully improved, the characteristics of green environmental protection and environmentalfriendliness are achieved, and wide applicability in the field of productive dust prevention and control is achieved.

The invention discloses a composite preparation for improving the culture survival rate of procambarus clarkii and an application method thereof. The composite preparation is mainly prepared from a component A and a component B, wherein the component A is prepared from 30-50% of bacillus and 50%-70% of aerobic denitrifying bacteria; the component B is prepared from 10%-15% of a quorum sensing quencher, 10%-15% of a synergist, 20%-30% of a flocculant, 10%-15% of a surfactant and 20%-30% of an oxygenating agent. The component B of the preparation is used firstly, and meanwhile the component A is used later after culture expansion. The preparation can improve the oxidative reduction potential at the bottom of a culture environment, reduce the propagation of cyanobacteria and microcystis aeruginosa in a culture water body, and decrease harmful flora in the bottom environment and harmful algae in the water body based on the principles of quorum sensing quenching, microbial ecological occupation, microbial matrix competition, microbial metabolite inhibition and the like by applying the component A and the component B in a synergistic sequence, so that the production of toxic substances such as microcystins, hydrogen sulfide, ammonia nitrogen and nitrite is basically avoided, and thus the survival rate of cultured procambarus clarkii is improved. The preparation has the advantages of quick action, strong effect sustainability and the like.

The invention belongs to the technical field of microbial culture and discloses an on-site rapid culture expansion method of aerobic denitrifying bacteria. The method comprises the following steps of(1) adding an aerobic denitrifying bacterial liquid into a first-stage culture tank, adding a culture medium, adding water for dilution, and carrying out oxygenation and stirring for culturing to obtain a bacterial liquid;(2) disinfecting two second-stage culture tanks, diluting each culture tank with water, and then performing oxygenation and stirring for culturing to obtain a bacterial liquid;(3) disinfecting two third-stage culture tanks, inoculating the bacterial liquid obtained in step (2) into one third-stage culture tank, filling the tank with a culture medium, adding water, and then performing oxygenation and stirring for culturing to obtain a bacterial liquid;(4) transferring half of the bacterial liquid obtained in step (3) into the other third-stage culture tank, adding a culture medium and water, and then performing oxygenation and stirring for culturing to obtain 2 tons of aerobic denitrifying bacterial liquid; (5) repeating steps (3) and (4) to obtain the ton-grade bacterial liquid according to demands. The culture expansion method has the advantages of simple operation, small investment and high practicability.

A production method of high-conversion and high-purity glucoamylase concentrate, mainly comprising: charging carbon dioxide, corn steep liquor, ammonium sulfate and water into the first-level seed tank according to the ratio; charging carbon dioxide into the second-level seed tank according to the ratio Raw material, bean cake powder, α-amylase, biotin and water; put carbon, bean cake powder, fed liquid ammonia, α-amylase, biotin and water in the fermenter according to the ratio; in the purification pretreatment tank Put sodium-based bentonite and acidified water in the medium according to the proportion; inoculate the Aspergillus niger strains into the first-level seed tank for cultivation according to the proportion; move the cultivated strains and raw materials into the second-level seed tank for expansion; The strains and raw materials are transferred to the fermenter for fermentation; next, the mature fermented liquid is put into the purification pretreatment tank for purification and pretreatment, followed by solid-liquid separation, concentration, fine filtration, sterilization and physical and chemical testing, and the finished enzyme is obtained after blending. The beneficial effect of the invention is that the purity of the glucoamylase system can be greatly improved, and the glucose conversion rate (DE value) can be obviously improved.

The invention discloses a multiple species inoculant for enhancing sediment nutrient. The multiple species inoculant comprises azotobacter chroococcum, bacillus megaterium and paenibacillus mucilaginosus; a volume ratio of bacteria solution of the azotobacter chroococcum to bacteria solution of the bacillus megaterium to bacteria solution of the paenibacillus mucilaginosus is 3:1:1. According to the invention, a segmented aerated culture method is adopted to carry out culture expansion, and the temperature is also changed with time of bacteria solution culture. The multiple species inoculant disclosed by the invention has the beneficial effects that the bacterial proportion of the multiple species inoculant can effectively enhance the sediment nutrient of an organic fertilizer, and the organic fertilizer can be decomposed to a great degree so as to enable crops to adsorb the organic fertilizer; compared with a multiple species inoculant which does not adopt segmented aerated culture, the multiple species inoculant adopting segmented aerated culture is more beneficial to bacterium breeding, enables a living bacteria count to be improved, and is beneficial for forming bacillus stearothermophilus spores; according to the multiple species inoculant disclosed by the invention, in different periods, different temperatures are adopted to culture bacillus, which enables the living bacteria count to be improved and is beneficial for forming spores.

The invention relates to methods of isolating and implanting mesenchymal stem cells for tissue repair or formation, without prior culture expansion of the mesenchymal stem cells. In particular, the invention relates to methods of isolating mesenchymal stem cells from bone marrow, for repairing or inducing formation of bone, without prior culture expansion of the mesenchymal stem cells. The invention further relates to an isolated, non-culurally expanded human adult mesenchymal stem cell population.

The present invention belongs to the technical field of preparation of probiotic microcapsules and particularly relates to a preparation method of a microcapsule of gastric acid-resistant bacillus licheniformis with high survival rates. The preparation method conducts an optimal control of the temperature, pH and culture medium during the culture expansion process, and the prepared polyelectrolyte microcapsule has a honeycomb core and an outer shell-like structure, and is preserved combining drying and vacuum-packaging. The produced bacillus licheniformis microcapsules are placed in a simulated gastric fluid, thus the bacillus licheniformis has high survival rates and high environmental tolerance. The product not only can be applied in the field of health-care food, but also can be applied in the field of medicine. The bacterial strains are probiotics and the carrier materials are natural polysaccharide macromolecules, thus the microcapsules are green, safe and environmentally friendly.

The invention discloses a system for microbial culture expanding and deep purification of a micro-pollution water body. The system comprises a grille, a micro-nano bubble machine, a microbial cultureexpander and a microbial enhanced treatment area composed of gravels, clay and submerged plants. Bottom sediment, especially the organic pollutants in floating sediment, can be effectively reduced, the ecosystem of a polluted water body can be quickly repaired and rebuilt, the self-purification capacity of the water body can be improved, and water quality can be improved. The polluted water body is lifted to the upstream of a riverbed, sequentially passes through the grille, the micro-nano bubble machine and the microbial culture expander and is introduced into an assimilative water body afterfiltration, oxygenation and rapid microbial culture expansion. Gravels, clay and submerged plants are arranged in the assimilative water body to strengthen a microbial treatment effect, create a natural habitat and improve the level of biodiversity. The system can achieve the circulation treatment of the polluted water body and has the advantages of high efficiency, safety, wide application range, strong anti-pollution and impact property, and the pollutants in a river channel are natural nutrient sources for microbial culture expansion.

The invention discloses a fertilizer additive prepared from a cell extracting solution and a preparation method of the fertilizer additive. The fertilizer additive is prepared from, by weight, acrylic acid, starch, N,N'-methylene bisacrylamide, chitin, ethephon, humic acid powder, moss, seaweed, corn flour, malt, gelatin, brown sugar, yeast powder, biogas slurry, dried potato powder, dried potato powder raw materials and a proper amount of water. Cells of moss and cells of the seaweed are cultured, organic materials can be decomposed through reproduction of the cells to generate organic matter decomposer, meanwhile, the organic matter decomposer can be subjected to culture expansion by itself, a certain amount of yeast is added, one part of cells which cannot be decomposed by the seaweed cells can be decomposed through the yeast, nutrients are provided, and finally the fermentation materials and the seaweed cells are crushed and extracted. On the one hand, organic acid ingredients beneficial to growth of crops generated after decomposition are utilized, and meanwhile intracellular products released after rupture of the cells of the moss and the seaweed cells also achieve a critical function on growth of the crops.

The invention discloses a nutritional bacterium agent, a preparation method and application of the nutritional bacterium agent in preparing bacterium fertilizers, belonging to the technical field of fertilizer preparation. The nutritional bacterium agent is characterized by comprising the following raw materials in parts by weight: 25-40 parts of spore bacteria, 20-35 parts of nitrifying bacteria,12-20 parts of yeast, 5-10 parts of photosynthetic bacteria, 6-12 parts of rhizobium, 5-9 parts of phosphobacteria and 4-8 parts of potassium bacteria. The nutritional bacterium agent is prepared bythe steps of activation and culture expansion and the like. After the nutritional bacterium agent disclosed by the invention is applied in bacterium fertilizers, the fast health growth of corns can bepromoted, and the contents of protein and vitamin in the corns can be increased.

HER2+ invasive breast cancer (IBC) patients with residual disease following neoadjuvant chemotherapy have an anti-HER2 Type 1 T helper (Th1) cell immune deficit and a significant risk of recurrent disease. It has been shown that anti-HER2 CD4+ T-cell responses incrementally decrease along the breast cancer continuum - a robust response in healthy donors and patients with benign disease, a depressed response in patients with HER2+ ductal carcinoma in situ, and a nearly absent response in patients with HER2+ IBC. This invention relates to a method of creating a microenvironment for culture expansion of T cells. The expanded T cells can be used for a variety of therapeutic and research purposes.

The invention relates to the technical field of cell engineering, in particular to a method for extracting, culturing and amplifying human adipose stem cells, comprising an extraction step of human adipose stem cells and a step of cultivating and amplifying human adipose stem cells; the extraction of human adipose stem cells is carried out through human fat digestive juice Realization, described human fat digestion liquid comprises: cholic acid, glycine, α1-antitrypsin, dimethyl sulfoxide, 0.9% normal saline, magnesium chloride, choline chloride, Trypsin-EDTA; The growth is realized through the adipose stem cell culture medium, and the adipose stem cell culture medium comprises: DMEM / F12 basal medium, thyroxine, insulin, glutamine, iron gluconate, sodium selenate, vitamin C, fibroblast growth factor, L-proline, choline chloride, penicillin; the method for extracting and culturing human adipose stem cells described in the present invention can significantly improve the quantity, survival rate and activity of the obtained stem cells.

A method of creating a microenvironment for culture expansion of T cells. The expanded T cells can be used for a variety of therapeutic and research purposes.