101results about How to "Increase culture density" patented technology

The invention discloses a blood serum-free culture medium suitable for the suspension culture of CHO cells with large scale and high density. The culture medium takes DMEM/F12 (v/v, 1:1) as a base culture medium, and adds the other substances such as insulin, putrescine, transferrin and microelement, etc. The blood serum-free culture medium has the advantages that: the culture medium can support the CHO cells to fast growth under the condition of suspension culture; the protein content is very low, and the chemical composition is basically clear; the cells are grown one by one by means of suspension, so that the living cell density and the cell living rate are higher than those of the serum culture and the similar commercialized serum-free culture; and the cost is lower.

The invention provides a method for preparing a rabies vaccine stock solution from serum-free Vero cells. The method comprises the following steps: culturing Vero cells with a serum-free culture medium to obtain a serum-free culture medium adaptive cell line; establishing a serum-free Vero cell seed bank by utilizing the serum-free culture medium adaptive cell strain; recovering, culturing, performing passage and amplifying cells of the Vero cell working seed bank by using the serum-free culture medium to serve as basal cells for bioreactor culture, and continuously perfusing to culture high-density Vero cells with the serum-free culture medium by applying a bioreactor and a microcarrier after cell expansion; inoculating virus seeds of a rabies vaccine virus strain working seed bank, performing bioreactor microcarrier serum-free culture, starting to continuously perfuse to obtain a virus fluid when the virus is amplified to peak to obtain titer of the virus fluid; and performing clarifying, ultrafiltration concentration, inactivation and purification treatment to obtain the serum-free human rabies vaccine stock solution.

The invention discloses a device for purifying flue gas by utilizing microalgae source photosynthetic microorganisms and a method thereof, which relate to environmental biological resource engineering. An LED microalgae source photosynthetic microorganism friendly illuminant of the system conducts light wave produced by an LED to the surfaces of the microalgae source photosynthetic microorganisms through light propagation media-optical fiber, and the microalgae source photosynthetic microorganisms utilize the photosynthesis to purify pollution gas such as CO2 in the flue gas dispersed in a microalgae source photosynthetic microorganism culture fluid containing growth factors so as to produce a large quantity of the microalgae source photosynthetic microorganisms as well as purify atmospheric pollutants simultaneously, wherein the microorganisms contains chlorophyll, protein and other useful organic substances. The device and the method recycle the pollutants, namely improve the efficiency of energy utilization, also eliminate the pollution, and have a green process with stronger practicability. The method has the advantages of less equipment investment, easy operation and high culture density of the microalgae source photosynthetic microorganisms in a reactor, and performs the resource utilization on the pollutants as well as purifies the pollutants simultaneously.

The invention relates to a process for preparing an avian influenza inactivated vaccine by full suspended culture cells and a product. The avian influenza inactivated vaccine product is prepared by the following steps: inoculating an avian influenza virus by the full suspended culture MDCK cells; clarifying, inactivating, degerming, concentrating and purifying and emulsifying the obtained avian influenza virus liquid. Compared with the prior art, the process provided by the invention is optimal and low in cost. Under the condition of guaranteeing the virus titer, the MDCK cell culture density can be greatly improved by the process provided by the invention, so that amplification on a large scale is easy to realize, and the avian influenza virus is proliferated in a great quantity. Compared with existing carrier suspended culture, the process provided by the invention is free from carrier intervention, so that the risk on pollution is greatly reduced. The process provided by the invention efficiently solves the problems of high pollution rate, low antigen output, long production period, insufficient SPF chicken embryo supply and the like of a chicken embryo method and is suitable for popularization and application of the reactor cell suspension culture technology in the avian influenza industry.

The invention relates to a cell culture module and a cell culture system. The cell culture module comprises a sealed body, a culture solution cavity and a cell cavity, a sealing stopping block and hollow fiber yarns, wherein the sealed body is communicated with the outer side through a culture solution inlet, a culture solution outlet, a cell sap inlet and a cell sap outlet; the culture solution cavity and the cell cavity are located in the body; the culture solution cavity comprises a culture solution cavity communicated with the culture solution inlet and a culture solution cavity communicated with the culture solution outlet, which are independent; the cell cavity is communicated with the cell sap inlet and the cell sap outlet; the sealing stopping block is used for separating to form the cell cavity and the culture solution cavity; the hollow fiber yarns are arranged on walls of the cell cavity and are provided with micro-pores which do not allow cells to pass; the hollow fiber yarns penetrate through the sealing stopping block; two ends of the hollow fiber yarns are communicated with the culture solution cavity located at a culture solution inlet end and the culture solution cavity located at a culture solution outlet end respectively. The cell culture system comprises a cell culture chip and can be used for culturing various cells.

The culture equipment of bait microalgae mainly is formed from microalgae culture bag, light source, air pipe, guide shell, oxygen-enriching pump and tourmaline, in which the microalgae culture bag is mounted on the base seat, and is positioned in the cylindrical steel wire ring, the bottom portion of the microalgae culture bag is equipped with algae liquor discharge outlet which is communicated with bait tank, and the top of the microalgae culture bag is equipped with water inlet pipe communicated with the cultured algae seed, prepared nutrient liquor and disinfected sea water, its interior is equipped with a guide shell, and the air pipe is positioned in the guide shell. Said equipment can implement all-weather high-density culture of bait microalgae, and its method is simple.

The invention relates to the field of veterinary biological products and particularly relates to a preparation method of a mink canine distemper virus live vaccine. The method comprises the following steps: inoculating a bioreactor with sensitive cells for vaccine preparation, and culturing by using a micro-carrier; after the sensitive cells are cultured by over 50% and grow into a dense single layer, inoculating the bioreactor with canine distemper virus for enrichment culture; harvesting the virus culture liquid and micro-carrier; performing freeze-thawing and removing the micro-carrier and cell debris to obtain virus liquid; and blending the virus liquid to obtain the vaccine. By improving the reaction conditions of each step and optimizing the production flow, the method provided by the invention realizes the technical effects of short production cycle, high virus titer, stable product quality, increased production efficiency and low side effect.

The invention discloses macro-element nutrient solution for large-scale culture of Chlorella salina. The nutrient solution is nutrient solution (1000x) prepared by using 9 to15 parts of urea, 1 to 3 parts of sodium dihydrogen phosphate, 0.3 to 0.5 part of ferric trichloride, 1.3 to 1.8 parts of EDTA-2Na and balance of water. The nutrient solution disclosed by the invention is used for the large-scale culture of Chlorella salina, has the advantages that the growth of Chlorella salina can be promoted, the culture density is improved, the culture efficiency is improved and the cell nutrient components of the obtained Chlorella salina are stable, and further has the characteristics of high culture density, long duration and low cost.

The invention relates to a method for suspension culture of Newcastle disease virus (NDV) by using a full-suspension passage cell line. The method comprises the following steps: inoculating the full-suspension EB66 cell line having a growth density of 8*10<6> cells/mL to 16*10<6> cells/mL with the NDV by using a two-stage culture method, continuously culturing after the virus is inoculated, and sampling every 12 hours so as to determine the EID50 of the virus; harvesting the virus when the titer of the virus is maximum, and storing the harvested virus so as to complete the culture of the NDV.The method adopts the duck embryonic stem cell EB66 passage cell line to culture the chicken NDV, thus effectively improving the titer and purity of the obtained virus, further enabling produced vaccines to have higher quality, and lowering the production cost.

The invention relates to tiny algae cultivating field, especially the high density cultivating method of Tama Alexander algae. In the initial stages, static laying and mildness whisking would be adopted to make the carbon source and other alimentation in the culture medium proliferate for a period of time. Taking aerate after the cell density reaching a certain extent, thus, injure of cell would be avoid. The gas could have a certain ratio CO2 to supply carbon source that the algae cell needs for growing. And a certain PH value would be sustained. At the initial stages, suitable N, P consistence would be supplied to make the cell grow fast. And at the late stages, N, P should be added to relief the lack of N, P, and achieve high cell density.

The invention discloses a solar energy internal light source microalga bioreactor. The bioreactor comprises a runway pool, a stirring impeller, an air feeder and a liquid feeder, and solar energy internal light source modules are arranged in the runway pool. In the invention, the advantages of an open photo-bioreactor and the advantages of a closed photo-bioreactor are fully combined through installing the solar energy internal light source modules having a simple structure and a low cost on the basis of the runway pool. The bioreactor disclosed in the invention, which allows the deep internal of an alga liquid to be illuminated, greatly increases the illumination surface area volume ratio of the alga liquid in a culture pool, improves the solar energy utilization rate of the photo-bioreactor and greatly increases the culture density of the open photo-bioreactor. The wall suspension of microalga appears on the external surfaces of the modules, the external surfaces of the modules are smooth and structured and have no protruding parts, and the installed modules can be dismounted at will, so automatic batch cleaning or manual cleaning is convenient.

The invention discloses a large-scale microalgae cultivation method, and belongs to the technical fields of microalgae cultivation and production. The method comprises the following steps: firstly, carrying out microalgae seed holding and enlarging cultivation in a laboratory, and then carrying out large-scale microalgae species solution cultivation in an outdoor light bioreactor; inoculating microalgae species solutions cultivated by using the light bioreactor into an outdoor runway culture pond, and carrying out large-scale microalgae cultivation in the runway culture pond; and finally obtaining microalgae biomass from the runway culture pond, and producing the microalgae biomass-based product. According to the method, through the combination of the outdoor light bioreactor and the runway culture pond, large-scale microalgae cultivation can be carried out; compared with the microalgae cultivated by a pure runway culture pond or a pure light bioreactor, the production cost is significantly reduced; and a whole set of complete process route is developed.

The invention discloses a method for continuously massively producing recombinant adenoviruses. The method comprises the following steps: 1, cell culture; 2, cell culture bag culture; 3, large-scale amplification of the adenoviruses; and 4, purification, wherein cell culture bags in step 2 and step 3 are placed on a shaking table, and culture in a thermotank is carried out; and the culture conditions of the cell culture bags are as follows: the temperature is 37 DEG C, the pH value is 7.2, the shaking speed is 10-20 rpm, and the angle of the shaking table is 7-9 DEG. The cell growth environment is improved, damages of ventilation to cells are reduced, and the dissolved oxygen effect is increased, so the problems of difficult digestion, separation and transfer of cells in traditional technologies are solved, thereby the enlargement culture problem and the continuous production problem of adherent cells are solved, and the problem of difficult freeze-thaw cracking of the cells is solved;and the adherent cells are cultured in the cell culture bags filled with a sheet carrier and the shaking bed, so the damages of a mechanical shearing force to the cells are reduced, and the growth density of the cells is improved, thereby the total cell yield of the cells is improved, and the yield of the adenoviruses is increased.

The invention relates to a high-density fermentation method and a product storage method of organophosphorus hydrolase recombinant strains, which belong to the technical field of environmental biotechnology. The invention optimizes the high-density culture conditions of organophosphorus hydrolase MPH recombinant strains Eshcherichia coliBL2l (DE3) pETMb, so that the optical density of fermentation fluid OD after 12 hours of fermentation reaches 29, and the enzyme activity reaches 18.5IU/ml. In addition, appropriate protective solution is added in the MPH production, so as to prolong the storage life of the enzyme, and the enzyme can also be mixed with a bio-surfactant, and simplify the use procedure of degrading enzyme. The invention can serve as an optimized formula and be applied to the production of fruits and vegetables detergent with enzyme, so that the invention plays a greater role in the pesticide degradation on the surface of fruits and vegetables. Through the storage scheme of the invention, the storage life of the enzyme can be prolonged to above 90 days under the storage conditions of liquid.

The invention discloses a recombinant plasmid and a preparation method thereof, and a preparation method and applications of a cell prepared by using the recombinant plasmid and capable of expressinghigh temperature-resistant alpha-amylase. Firstly, a recombinant plasmid containing a P43 promoter and lacO and lacI genes is prepared; then the high temperature-resistant alpha-amylase gene is connected to the recombinant plasmid, the plasmid contains a bacillus subtilis strong promoter P43 so as to greatly improve the yield of high temperature-resistant alpha-amylase, meanwhile, the sequences ofthe lacO and lacI genes contained in the plasmid can be subjected to induced expression by using IPTG, and the induced expression is faster and more targeted than microbial mutagenesis breeding; andthen a receptor cell and a vector containing the high temperature-resistant alpha-amylase gene are subjected to electric shock transformation so as to obtain the cells capable of expressing the high temperature-resistant alpha-amylase. When bacillus subtilis is used as the receptor cell, the gene information is clear, the production speed is high, the culture density is high, and the yield and enzyme activity of alpha-amylase can be improved.

According to the invention, nutrient substance used for the growth of herba cistanche cells is widely selected through the experiment means, and the formula of the invention is determined according to the self metabolic properties and practical use effects of the herba cistanche cells. According to the formula, ammonium nitrate and potassium nitrate are taken as nitrogen sources, monopotassium phosphate is taken as phosphorus source, under the basic nutritional ingredients of the MS culture medium, the plant hormones including 2.4-D, 6-BA and NAA are added for the purpose of improving metabolites, meanwhile, the inducers including yeast extract and chitosan are added, through the above improvement, the growth rate of the herba cistanche cells is greatly improved, and the secretion amount of phenylethanoid glycosides is greatly increased. On the basis, according to the accidental discoveries, by adding the traditional Chinese medicinal materials with specific varieties and ingredients, the cell culture density can be improved. Based on the above beneficial discoveries, through the experiment means, the traditional Chinese medicinal materials used for treating the human diseases, such as herba tadehagis triquetri, herba saxifragae stoloniferae, herba ranunculi chinensis and herba abri are selected, and the obtained culture medium obtains the outstanding culture effects.

The invention provides a ULDPE-containing multi-layer co-extrusion disposable biotechnological bag membrane material and a preparation method thereof. A nylon/ethylene-vinyl alcohol copolymer/nylon three-layer composite structure is adopted to prepare a gas barrier layer, ULDPE is used as a liquid contact layer, and an optimal heat sealing layer, an optimal bonding layer and an appropriate thickness are selected, so that the prepared disposable biological process bag has good gas barrier property, water resistance, strength, bending resistance, puncture resistance, excellent heat sealability and transparency, has good biocompatibility, high cell culture density and better cell culture effect, is simple in preparation process, simple and convenient to operate and high in efficiency, and can be used for preparing disposable bioreactor bags, liquid storage bags, stirring bags, weighing bags, feeding bags and the like.

The invention provides a Japanese encephalitis virus suspension culture method. The technical scheme is developed on the basis of an animal cell microcarrier culture technology. As a microcarrier provides a larger surface area for the growth and reproduction of a monolayer adherent cell, a homogenic suspension culture system is provided for cell growth, the microcarrier is utilized to serve as physical support for cell colonization, and together with optimization of culture conditions, the cell and virus culture densities are improved. The Japanese encephalitis virus suspension culture method realizes large-scale culture of the Japanese encephalitis virus and provides lots of high-quality virus antigens. Moreover, the shortcomings that traditional spinner cultivation animal primary cell production is low in single-batch yield, great in batch difference, unstable in product quality and high in production cost and pollution probability are overcome. Besides, the Japanese encephalitis virus suspension culture method can achieve continuous culture, is small in land occupation, large in production scale and small in cell damage, and the shortcomings that a traditional process is large in land occupation, discontinuous in culture and high in production cost are overcome.

The invention discloses a bioreactor, a stirring paddle thereof and a method for cultivating TIL cells through the bioreactor. The stirring paddle is wrapped by a material for reducing shear force. The bioreactor comprises the stirring paddle wrapped by the material for reducing shear force, and TIL cells are cultivated through the bioreactor. As TIL cells are cultivated through the bioreactor, and the stirring paddle is wrapped by the material capable of reducing shear force, damage of shear force to cells can be reduced, and mass amplification of cells is facilitated. In the cultivating process, stirring continues, a dynamic system is provided, cells can conduct gas exchanging, the even cultivating environment can be provided for cells, and mass amplification of cells is facilitated. By means of the method, mass amplification of TIL cells can be conducted, the expression quantity of surface antigens of obtained TIL cells is high, and tumor cell killing activity is high.

The invention relates to a method for preparing an antigen protein for detecting a diphtheria antibody. The method is characterized by comprising the steps as follows: S1: infecting a baculovirus expression vector carrying a CRM197 protein expression cassette into insect cells so as to obtain infected insect cells; S2: carrying out cultivation and proliferation on the infected insect cells; and S3: carrying out crushing on the proliferated insect cells, and extracting the antigen protein in the insect cells. The invention further relates to an antigen protein prepared by the method and furtherrelates to a kit comprising the antigen protein. An expressed CRM197 protein is safe, non-toxic, high in yield and easy to separate and purify, meanwhile, has very high affinity with the diphtheria antibody, and is good in specificity.

The invention provides elicitor composition for producing paclitaxel from Taxus chinensis cells. According to the elicitor composition, components including phenylanine, sodium acetate, methyl jasmonate, salicylic acid, silver nitrate, arachidonic acid, EDTA (ethylene diamine tetraacetic acid), sodium citrate, calcium chloride, boric acid and the like are selected preferably, the dosage and ratio of the components are designed reasonably, and the using method of the elicitor composition is that the composition is directly added to a cell culture fluid at the initial culture stage or after cells are in the stable culture stage. Experiments find that after addition of the elicitor composition, although the cell culture density cannot be directly increased, the yield of paclitaxel in the same biomass of cell culture liquid is increased significantly, which indicates that the elicitor composition has a certain influence on the metabolism characteristic of the Taxus chinensis cells. The outstanding technical effect is realized by innovative technical means, and the present status of mass production of the paclitaxel is expected to be improved comprehensively by the aid of the elicitor composition.