135results about How to "Increase multiplier" patented technology

The invention discloses a culture medium for tissue culture of pedicel buds of oncidium hybridum and a tissue cultured seedling propagating method. The culture medium is mainly characterized in that: the combination of concentrations of macroelement components in the culture medium does not exist in the conventional culture medium for the tissue culture of the oncidium hybridum. The method for propagating tissue cultured seedlings of the oncidium hybridum comprises the following steps: utilizing the culture medium to induce the pedicel buds to generate callus and differentiation of protocorm; after the protocorm sprouts and tissue cultured seedlings provided with fake corms are formed, transferring to a rooting medium, and inducing to root; transplanting the rooted tissue cultured seedlings into the medium by a conventional tissue cultured seedling transplanting method. By adopting the technical scheme provided by the invention, the pedicel buds of the hybrid variety of the oncidium hybridum are taken as explants, young plants of the oncidium hybridum are quickly propagated in a way of differentiating the protocorm, and the speed of propagating the young plants of the oncidium hybridum is improved.

The invention relates to the technical field of industrialized seedling production and in particular relates to a blueberry tissue culture propagation and ex-vitro rooting method. The method mainly aims to solve the technical problems that rapid industrialized blueberry industrialized seedling production is restrained due to low rooting rate, long rooting period, complex rooting program, high rooting cost and the like during in-vitro blueberry rooting. The method comprises the following steps: selecting and sterilizing an explant, inducing lateral bud germination, performing multiplication culture, and finally performing ex-vitro rooting, wherein in the culture medium for inducing lateral bud germination, the components of the culture medium comprises an improved WPM basic culture medium, 0.8-2mg / L of ZT, 25-35mg / L of white sugar and 5-10g / L of agar; the pH value is regulated to be 5.4; according to the improved WPM basic culture medium, the original potassium sulfate and calcium chloride are replaced by calcium nitrate hydrate and potassium nitrate.

The invention discloses a preparation method of antitumor adoptive immune cells. The method comprises the following steps of: sampling and separating a single peripheral blood karyocyte; and adding a cell factor for inducting and culturing to obtain a cytokine induced kill cell (CIK), wherein the cell factor comprises CD28. In a CIK induced system, the cell factor CD28 is induced, so that the inducing effect is enhanced, and the cell proliferation multiple is increased simultaneously.

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.

A motor control apparatus provided with an inverter for successively commutating the current to a motor using a PWM signal; a PWM signal generating device for generating the PWM signal using a carrier signal; a rotational state quantity sensor for detecting a rotational state quantity; a phase difference detecting device for detecting the phase difference between the carrier signal and the rotational period based on the rotational state quantity; a frequency setting device for setting a frequency of the carrier signal to a value in accordance with a multiplier for one period in terms of electrical angle of the rotational period of the motor, when the rotational frequency is equal to or greater than a specified frequency and the phase difference is equal to or less than a specified value; and a synchronizing device for synchronizing a control period of the carrier signal to the rotational period.

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.

The invention discloses a proliferation medium for Sierra cowberry blueberry tissue culture. 1L of the proliferation medium comprises modified WPM (woody plant medium), 0.5mg of NAA (Naphthalene Acetic Acid), 0.2mg of GA, 2-3mg of zeatin (ZT), 30g of saccharose, and 7g of agar, and has pH value of 5.2. 1L of the modified WPM contains macroelements, iron salts, microelements and organic substances, wherein the macroelements comprise 893mg of potassium nitrate, 119mg of ammonium sulfate, 370mg of magnesium sulfate, 278mg of calcium nitrate tetrahydrate, and 170mg of monopotassium phosphate; the ferric salts comprise 74.6mg of ethylene diamine tetraacetic acid, and 55.6mg of ferrous sulfate; the microelements comprise 22.3mg of manganese sulfate, 8.6mg of zinc sulfate, 0.025mg of copper sulfate, 0.25mg of sodium molybdate, 6.2mg of boric acid, 0.415mg of potassium iodide; and the organic substances comprise 0.5mg of niacin, 0.5mg of thiamine hydrochloride, 0.5mg of pyridoxine hydrochloride, 100mg of inositol and 2mg of glycine. The proliferation rate can reach more than 12 after the medium is used for 30 days.

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 tissue culture and rapid propagation method of hemionitis arifolia seedlings by virtue of a green spherical body manner. Mediums used in the tissue culture and rapid propagation method comprise an induction medium, a propagation medium and a differential medium, wherein the induction medium is prepared from 0.4-1.0mg / L 1 / 6MS-1 / 4MS and TDZ, 0.1-0.3mg / L NAA and 400-800mg / L casein hydrolysate; the propagation medium is prepared from 0.1-0.3mg / L 1 / 6MS-1 / 4MS and TDZ, 0.5-1.0mg / L 6-BA, 0.1-0.3mg / L NAA and 400-800mg / L casein hydrolysate; the differential medium is prepared from 1 / 4MS-1 / 2MS and 3-6g / L active carbon; cane sugar and agar are added into all the mediums; the pH value of each medium is 5.80. The induction rate of the green spherical body is greater than 88%; the proliferation time is 11-12.5; the differentiation rate is greater than 98%; 1.5 million to 2 million of seedlings can be propagated from one green spherical body in one year; the survival rate s greater than 95%.

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 relates to a tissue culture seedling culturing method for fir wood of excellent clonal 11C. The method comprises the following steps:(1) the explant of the fir wood of the excellent clonal 11C is innoculated in an inducing culture medium, the inducing culture of an indefinite bud is performed, and the indefinite bud is obtained; (2) the indefinite bud is innoculated into a subculture proliferation culture medium, the subculture proliferation culture is performed, and a subcultured seedling is obtained; (3) the subcultured seedling is innoculated into a rooting culture medium, inducing rooting culture is performed, and a rooting seedling is obtained. The tissue culture seedling culturing method for the fir wood of the excellent clonal 11C, disclosed by the invention, has the advantages of high inductivity, high proliferation rate, high effective seedling rate and high rooting rate.

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 discloses an immune cell culture medium. The culture medium is prepared from the following components: anhydrous calcium chloride and other multiple inorganic salts, L-leucine and othermultiple amino acids, interleukin 2, interleukin 4 and the like. The immune cell culture medium can be used for remarkably increasing the proliferation rate of cells, greatly prolonging days for culturing cells and effectively keeping expression of surface markers when being used for culturing CAR-T cells, and has the advantages of long preservation period, easy preparation and the like.

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.

A disclosed dendrobium officinale Kimura et Migo tissue culture batch production method through one-step seedling formation comprises the following steps: (1) breeding protocorm, namely, selecting a terminal bud or an axillalry bud of an excellent dendrobium officinale Kimura et Migo plant as an explant, sterilizing the explant, putting in a MS agar solid medium with 6-BA concentration of 0.1-3.0 mg / L and KT concentration of 0.2-1.0 mg / L and IAA concentration of 0.1-1.0 mg / L and NAA concentration of 0.2-2.0 mg / L and chlormequat concentration of 0.01-0.5 mg / L, inducing protocorm and obtaining a robust bud through differentiation; (2) performing one-step seedling formation batch production, namely, directly transferring protocorm with the bud into an MS agar solid medium with 6-BA concentration of 0.2-1.0 mg / L and IAA concentration of 0.2-0.5 mg / L and NAA concentration of 0.1-2.0 mg / L and chlormequat concentration of 0.5-2.0 mg / L, and performing propagation, differentiation, seedling strengthening and rooting; and (3) transplanting the test-tube seedling. The provided dendrobium officinale Kimura et Migo high-efficiency low-consumption tissue culture method is short in breeding period and low in cost, is extremely suitable for industrialized production, and reaches industrialization level.

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 the technical filed of industrialized seedling production, in particular to a method for tissue culture and propagation expansion of iris. The technical problems of slow propagation expansion, long rooting period, complicated rooting procedure, high production cost and the like which restrict rapid industrialized seedling production of the iris are solved. The method comprises the steps that first an explant is selected and disinfected, then tender shoots are induced to germinate, then proliferation culture and rooting culture are performed, and finally domestication and transplanting are performed. The medium which induces tender shoot germination is prepared form the components: modified MS basic medium, 1-2mg / L of 6-BA, 0.3-0.5mg / L of IBA, 25-35mg / L of sugar and5g / L of agar, pH is adjusted to 5.8, and the modified WPM basic culture replaces original potassium nitrate and calcium chloride with calcium nitrate hydrate and potassium sulfate; and the root dripping treatment is carried out again by domestication and transplanting.

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 belongs to the field of cytobiology and particularly relates to a culture medium and method for culturing DCs. The culture medium for culturing the DCs comprises a first component, a second component and a third component. The first component is an RPMI1640 culture medium containing bovine serum albumin, interleukin 4, GM-CSF and stem cell factors (SCF). The second component is an RPMI1640 culture medium containing bovine serum albumin, interleukin 14, interleukin 2, interleukin 4 and GM-CSF. The third component is an RPMI1640 culture medium containing bovine serum albumin, CD40L, GM-CSF and interleukin 4. The result shows that the antigen presentation capacity of the DCs cultured by the culture medium is higher, the sensitization capacity to T cells is higher, and the killing capacity to tumors is greatly improved. Use of serum is avoided, and the risk of pathogenic microorganisms is relieved.

The invention provides a tissue culture method of Stachys siebolidii Miq. According to the method, a propagation system of commercial test-tube seedlings is obtained through the steps of induction, propagation expanding, seedling strengthening, rooting and the like of Stachys siebolidii Miq immature stem segments in an in-vitro condition, thereby shortening the propagation cycle, increasing the proliferation rate, making the seedlings consistent in grow state, excellent in quality and not restricted by seasons, achieving anniversary production, and achieving the propagation of high-quality commercial Stachys siebolidii Miq seedlings through a tissue culture mode.