135results about How to "Increase multiplier" patented technology

The invention provides an NK cell serum-free culture medium. The culture medium comprises two independently packaged components which are used in different stages of cell culture, so the NK cell culture process is divided into an induction stage and a proliferation process, the culture effect is improved, and the cell induction and culture process is simplified, that is, other extra cell induction factors do not need to be added in the culture process. Polyinosinic cells are added into the NK cell serum-free culture medium and are more suitable for NK cell efficient culture, and the proliferation rate is increased.

The invention relates to an efficient CIK cell preparation and detection method and aims at solving the problems of few proliferation times, unstable preparation process and the like of the prior art. The efficient CIK cell preparation and detection method comprises the following steps that sterile acquisition is conducted peripheral blood of a healthy person or a patient according to a lymphocyte absolute value; whole blood testing and blood sample preservation are performed; separation and inactivation of blood plasma are performed; a density gradient centrifugation method is adopted, namely a single mononuclear cell is separated out by using lymphocyte separation liquid Ficoll; in the CIK cell induction process, a factor 1 and a factor 2 are added successively, collection can be performed only through serum-free induced cell multiplication culture for about 2-3 weeks, and a CIK cell preparation is prepared. The method has the advantages of being small in blood collecting quantity, stable in preparation process, simple and convenient to operate and high in cell proliferation times and cytotoxic activity.

The invention belongs to the technical field of immune cells in-vitro culture, and discloses a method for culturing cord blood lymphocyte DC-CIK. According to the invention, cord blood is taken as a processing object, and is subjected to separation of cord blood mononuclear cells, isolated culture of DC cell, induction culture of CIK cell, and co-culture of DC and CIK to prepare the DO-DIK cell, CTLA-4mAb and PD-1mAb are added, and the maturated DO-DIK cell is subjected to centrifugal collection to obtain the product. By using CD3mAb coating culture bottle, continuous stimulation is carried out at an early stage of culture, cytokine usage is reduced, effector cell population activation efficiency and amplification efficiency are increased, and the culture cost is reduced.

An airlift bioreactor for culturing the adventitious root of red sage is composed of an air compressor, an air buffering tank, the first and the second air filters, an air distributor, and a main reactor body consisting of cylindrical body, truncated conic body and spherical body. It can supply air at constant flow and variable air speed, so improving the oxygen supply in reactor.

The invention discloses an establishment method of an efficient regeneration system of mniochloa abersend shoots. The establishment method comprises the following steps: (1) carrying out induction culture on a bud callus; (2) carrying out enrichment culture on the callus; (3) carrying out differentiation culture on the callus; (4) carrying out rooting culture of test-tube plantlets; and (5) hardening and transplanting regenerated plants. According to the invention, a plant tissue culture method taking the shoots of herbal bamboo as an explant is provided, which is a useful attempt for establishing the regeneration system by callus induction with the shoots of herbal bamboo as the explants; with the method, the callus can be efficiently induced through the explants, thus the efficient regeneration system is established, the research field of the bamboo is expanded, and good foundation is laid for the genetic transformation research of the bamboo; and moreover, the method is easy and feasible, easy to popularize, loose in implementation conditions, and remarkable in effect.

The invention discloses a method capable of effectively inhibiting browning for the multiplication culture of buckwheat callus, which is characterized in that before the multiplication culture, the buckwheat callus is pre-cultured, and citric acid and L-cysteine are added into a multiplication culture medium. Therefore, the browning rate of the buckwheat callus during the multiplication culture is effectively inhibited, the multiplication times of the buckwheat callus is increased, and a new method is provided for greatly and quickly producing flavonoids (rutin).

The invention belongs to the technical field of tissue culture production and relates to a tissue culture fast propagation method for garcinia paucinervis.The method includes the following steps of 1, disinfection of an explant; 2, callus culture; 3, callus bud differentiation; 4, bud rooting.The formula of a root inducing medium is MS + 6-8g/L agar + 25-29g/L saccharose + 0.8-2.5mg/L indoleacetic acid + 0.5-1.5mg/L kinetin + 0.8-1.5mg/L activated carbon + 0.1-0.5mg/L metaxamin + 0.5-1.5mg/L compound sodium nitrophenolate + 0.3-0.8mg/L vitamin D2 + 0.15-0.35mg/L microcystin.The tissue culture fast propagation method has the advantages that propagation cost can be reduced, the growth period is short, the propagation rate is high, management is convenient, industrial production and automatic monitoring are benefited, and manpower, material resources and land for field crops are greatly saved.

The invention belongs to the field of biological tissue engineering cytoskeleton materials, and relates to a cross-linked microsphere as well as a preparation method and application thereof. Accordingto the method, chitosan-gelatin physical mixed microspheres are prepared in a spray freeze drying mode, and then amino and carboxyl in the physical mixed microspheres are subjected to chemical crosslinking, so that crosslinked microspheres are prepared. The problem that the microsphere structure is unstable due to a single spray freezing mode is solved; meanwhile, the cell adhesion characteristicof the microsphere can be improved by combining chitosan and gelatin, cell culture efficiency can be improved by using the microsphere as a microcarrier, and two-dimensional static culture and three-dimensional dynamic culture can be achieved. The chitosan and gelatin raw materials are low in cost, the large-scale production cost of the physically mixed microspheres and the chemically cross-linked microspheres is low, toxic substance residues are avoided, large-scale cell culture can be conducted, and great application prospects are achieved in the aspect of in-vivo injection.

The invention discloses autologous NK cells and a culture method and application thereof. The culture method of the autologous NK cells comprises the following steps of collecting peripheral blood mononuclear cells of a patient, combining the mononuclear cells, anti-HER2 monoclonal antibodies or HER2 monoclonal antibodies and fibronectin for use and adding cell factors into a serum-free medium for induction and multiplication culture. According to the culture method of the autologous NK cells, the anti-HER2 monoclonal antibodies and the fibronectin are adopted for improving the irritating activity of irritation for inducing an individual cell; the cell factors are adopted for inducing the autologous NK cells, the synergistic effect is generated on multiplication of the NK cells by means of combined application of the cell factors, the killing activity of the autologous NK cells is improved, and toxic and side effects caused by applying a large dosage of single factors are reduced. In addition, by means of the culture method, the culture time is shortened, the culture cost is reduced, and the purity of the autologous NK cells is guaranteed.

The invention belongs to the field of biological treatment of tumors and relates to an immune cell culture method capable of simultaneously activating CD4 <+>and CD8<+>T cells. The immune cell culture method comprises the steps of placing single nuclear cells which are well separated in a culture solution containing rhIL-12 and IL-4 ligands and an IL-10 ligand, culturing for 8-12h, then adding levamisole, continuously culturing for 8-12h, adding Anti-CD3McAb and rhIL-2, continuously culturing for 2-3 days, and then alternately adding into the culture medium containing the rhIL-2 and the culture medium containing the Anti-CD3McAb and the rhIL-2, wherein the intermediate culture time is 2-3 days. The scheme for preparing a cell factor compound is as follows: collecting cell supernatant liquid within 3-14 days after the beginning of culture, performing low-temperature ultracentrifugation, performing ultrafiltration by an ultrafiltration centrifugal tube, and performing microfiltration by a microprous filtration membrane, wherein filtrate is a natural cell factor mixture. By adopting the culture method provided by the invention, sustained proliferation of cells can be kept for 30 days. NCM contains a variety of cell factors and can stimulate immune cells of a human body to play autoimmune anti-tumor activity.

The invention discloses a method for improving the tissue culture and rapid propagation proliferation rate of Fraxinus velutina. The method is characterized in that a WPM medium is used as a basic medium in the rapid propagation proliferation culture process of tissue culture plants, and four plant growth regulators with different concentrations are proportioned to greatly improve the propagation rate of Fraxinus velutina tissue culture plants; and a 1/2 WPM medium with harving macroelements is used as a basic medium, a combination of two auxins, a cytokinin and low sugar is used to induce rooting of the tissue culture plants in order to obtain complete plants. The method effectively solves the problem of low proliferation rate of Fraxinus velutina. Results of experiment statistics show that the proliferation rate of test tube plants improves to above 10 times from original highest 3.59 times, the proliferation coefficient improves 2-3 times, and the rapid propagation proliferation purpose of the Fraxinus velutina is realized. A large amount of seedlings can be obtained within a short time by using the method, and can fully meet the forestation need of a large area of saline-alkali soil.

The invention belongs to the technical field of immune cell in-vitro culture, and particularly discloses a culture method of umbilical cord blood lymphocyte CIK. The method comprises the following steps: by using umbilical cord blood as a treatment object, separating the umbilical cord blood mononuclear cells, purifying and inducing the CIK cells, culturing and amplifying the CIK cells, and finally, centrifuging to collect the mature umbilical cord blood CIK cells, thereby obtaining the product. The IFN-gamma and alpha-galactose ceramide are added to activate the CIK cells, and CD3 monoclonalantibodies, IL-1 alpha, IL-2 and IL-1 beta are added in the early culture stage to perform continuous stimulation, thereby saving the coating time and enhancing the activation efficiency and amplification efficiency of effector cell groups.

The invention relates to a method for plant propagation, in particular to a method for directly inducing and quickly propagating a bog bilberry root nodule in a test tube. The method is characterized by comprising the following steps of: (1) selecting a tiller bud of a bog bilberry root as an explant, wherein a proper nutrient medium for inducing a bud root nodule is 1/4MS+IAA0.80mg.L-1+IBA0.04mg.L-1+GA30.15mg.L-1; and (2) selecting a stem node growing the root nodule already of a reproducing plant as a material to perform quick propagation in the test tube by a propagation method, cutting a seedling which contains the root nodule in the test tube from the upper part of the root and leaving 1 or 2 leaves on the cut part, cutting the cut twig into sections which have a leaf respectively, grafting the sections into the optimized root nodule induction nutrient medium to allow auxiliary buds to sprout and grow and culturing the root nodule to regenerate the root nodule. The method shortens a successive multiplication cycle, improves propagation folds, is simple, economical, practical and strongly operable, achieves the quick propagation aim and can be used for the industrial breeding of the bog bilberry having the root nodule.

The invention relates to a culture medium and culture method for inducing callus differentiation through peony leaves. The method comprises the following steps: in late March, selecting tender peony leaves; after sterilizing the leaves, inoculating the sterilized leaves to an induction medium, wherein the temperature is 25+/-2 DEG C; culturing in dark for 2 months; inducing to form callus; transferring the callus to a differential medium, wherein the temperature is 25+/-2 DEG C; the illumination time is 12 h.d<-1>; the illumination intensity is 1200+/-1001x; culturing for 2 months to obtain adventitious buds. According to the method, the callus initiation rate of peony leaves can reach up to 98.60%, the vitrification ratio is 0%, and the differentiation ratio of the adventitious buds can reach 58.99%; each callus block can form 4-10 adventitious buds, so that the technical problem that the ratios of adventitious bud differentiation is low when peony leaves are adopted as explants to induce callus is solved; the method provides technical support for carrying out peony gene engineering, and is wide in market prospect.

The invention discloses a method for promoting the polysaccharide accumulation of hairy roots of beautiful millettia roots. The method comprises: inoculating hairy roots of beautiful millettia roots into a culture medium, and carrying out multiplication culture on a constant-temperature shaking table for at least 30 days, wherein the liquid culture medium contains 100-300mu m mol/L methyl jasmonate. According to the method for promoting the polysaccharide accumulation of hairy roots of beautiful millettia roots, disclosed by the invention, the hairy roots of the beautiful millettia roots are subjected to amplification culture under a specific condition, so that the content of beautiful millettia root polysaccharide in the hairy roots of the beautiful millettia roots is increased, and a new approach is provided for obtaining beautiful millettia root polysaccharide.

The invention discloses a DC-CIK cell culture medium, a DC-CIK cell culture method and application, and relates to the technical field of cell culture. The DC-CIK cell culture medium comprises a DC culture medium containing specific components, a DC maturation factor, a CIK promoter, a CIK bottle enlargement culture medium, a CIK separate culture medium, a coating liquid and a basic culture medium, and all the components have different effects through different compositions. When the DC-CIK cell culture medium is used for culturing the DC-CIK cells, the number of the obtained DC-CIK cells is substantially increased and can reach (1.05-1.56) * 1011/mL, the survival rate is kept at 96.6%-99.7%, meanwhile, the CD3+, CD4+ and CD5+ of the DC-CIK cells can reach 68%.

The invention discloses a novel NKT cell culture method. The method comprises NKT primary cell culture and NKT cell subculture. The NKT primary cell culture comprises inoculating a serum-free medium with peripheral blood mononuclear cells. The NKT cell subculture comprises adding a promoter humanized CD3 monoclonal antibody, a humanized CD28 monoclonal antibody and IL-2 into the serum-free medium in the first day of the NKT cell subculture and adding amplification factors IL-2 and IL-7 into the later amplified solution. The NKT cells cultured by the novel NKT cell culture method have a good cell state, a fast propagation speed, high proliferation multiple, high purity and killing effects obviously better than those of NKT cells obtained by the existing culture method.

The invention discloses a CIK (cytokine induced killer) cell and a culture method and an application thereof. The culture method of the CIK cell comprises the following steps of collecting a peripheral blood mononuclear cell of a patient, adding the peripheral blood mononuclear cell, an anti-CD3 antibody, an anti-CD28 antibody and cell factors into a serum-free medium, and performing induced culture. The culture method of the CIK cell has the advantages that the stimulation activity for stimulating and inducing the mononuclear cell is improved under the synergetic action of the anti-CD28 antibody and the anti-CD3 antibody; the CIK cell is jointly induced by the cell factors IL-2 and IL-21, so that the joint application of the cell factors can generate the synergetic action on the amplification of lymphocyte, the killing activity of the CIK cell is improved, and the toxic or side effect caused by the application of multiple single factors is reduced; the culture time is shortened, the culture cost is reduced, and the purity of the CIK cell is guaranteed.