20results about How to "Improve proliferation efficiency" patented technology

The invention discloses a serum-free culture system and culture method of NK cells. The invention firstly discloses a serum-free culture system of the NK cells, wherein the serum-free culture system comprises a coating solution, an amplification culture medium, an activation culture medium and autologous plasma; the invention further discloses a culture method for NK cells by using the serum-free culture system. The method for culturing the NK cells by using the serum-free culture system disclosed by the invention reduces the culture cost of the NK cells, can simply, quickly, efficiently and safely obtain a large number of NK cells with high activity, high purity and strong killing ability in vitro, provides a better cell source for clinical application of the NK cells, and promotes further development of an NK cell treatment technology.

The invention relates to a cell culture microcarrier as well as a preparation method and application thereof. The cell culture microcarrier comprises a polymer porous microsphere with an inside-outside through pore channel structure. The cell culture micro-carrier comprises a biodegradable micro-carrier and also comprises a non-degradable micro-carrier, so that the growth of cells in the carrier is facilitated, and the mass transfer of nutrient substances and metabolites is also facilitated, thereby promoting the growth of the cells, enabling the cultured cells to keep good activity and function, and being high in proliferation efficiency; the method is suitable for three-dimensional culture of somatic cells, stem cells, tumor cells and the like.

The invention provides a preparation method of a PSR.Syn.CD19. CD38 CAR-T cell as well as the cell and application of the PSR.Syn.CD19. CD38 CAR-T cell, the PSR.Syn.CD19. CD38 CAR-T cell is characterized in that after the CAR-T cell and a CD19 antigen positive tumor cell are co-incubated, effector molecule anti-CD38-CAR can be induced to express, and when the tumor cell simultaneously expresses CD19 and CD38 antigens, the CAR-T cell capable of recognizing and killing tumors can be identified. The CAR-T cell provided by the invention can efficiently and specifically kill tumor cells (CD19 + CD38 +), has a persistent killing effect, and has a safety switch capable of starting a killing function. Besides, the tumor can be specifically killed, the cell proliferation rate is high, the anti-tumor function can be improved through a large amount of secreted IL-2, IFN-gamma, TNF-alpha and Granzyme B, and the application safety of the cell is greatly improved.

The invention discloses a cord blood-derived hematopoietic stem cell as well as an amplification method and application thereof. The method comprises the step of culturing umbilical cord blood mononuclear cells by using a culture medium containing thymus active polypeptide, stem cell growth factors and heat shock protein. According to the method for amplifying the umbilical cord blood-derived hematopoietic stem cells, the proliferation efficiency of umbilical cord blood mononuclear cells can be remarkably improved, and the amplification capacity of umbilical cord blood immune cells (NK cells) is improved by more than 5 times compared with that of a traditional method.

The invention discloses an apple rootstock tissue culture seedling raising method, and relates to the technical field of tissue culture. The method comprises the following steps: (1) washing annual robust branches with tender shoots in a dormancy period with running water, soaking with soapy water, cleaning, obliquely shearing into stem segments, soaking with alcohol, washing, sterilizing, washing, absorbing moisture, and shearing off wounds to obtain pretreated explants; (2) inoculating the pretreated explant into a differential culture medium, and culturing to obtain an adventitious bud; (3) transferring the adventitious buds into a proliferation culture medium, and performing dark culture and light culture to obtain adventitious bud clusters; (4) inoculating the adventitious bud clusters into a rooting culture medium, and culturing to obtain tissue culture seedlings; and (5) hardening the tissue culture seedlings, washing off the culture medium, disinfecting, putting into a culture medium, culturing to obtain apple seedlings, and transplanting to outdoor soil. According to the method, the seedlings capable of being transplanted outdoors can be rapidly obtained, and the problems of infectious microbe pollution, low survival rate, poor rooting effect and the like in existing tissue culture seedling culture are effectively solved.

The invention discloses an apple rootstock tissue culture seedling raising method, and relates to the technical field of tissue culture. The method comprises the following steps: (1) washing annual robust branches with tender shoots in a dormancy period with running water, soaking with soapy water, cleaning, obliquely shearing into stem segments, soaking with alcohol, washing, sterilizing, washing, absorbing moisture, and shearing off wounds to obtain pretreated explants; (2) inoculating the pretreated explant into a differential culture medium, and culturing to obtain an adventitious bud; (3) transferring the adventitious buds into a proliferation culture medium, and performing dark culture and light culture to obtain adventitious bud clusters; (4) inoculating the adventitious bud clusters into a rooting culture medium, and culturing to obtain tissue culture seedlings; and (5) hardening the tissue culture seedlings, washing off the culture medium, disinfecting, putting into a culture medium, culturing to obtain apple seedlings, and transplanting to outdoor soil. According to the method, the seedlings capable of being transplanted outdoors can be rapidly obtained, and the problems of infectious microbe pollution, low survival rate, poor rooting effect and the like in existing tissue culture seedling culture are effectively solved.

The invention discloses a serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and application thereof. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition is prepared from the following components in parts by mass: 15 to 25 parts of platelet-derived growth factor-BB; 15 to 25 parts of basic fibroblast growth factors; 8 to 12 parts of transforming growth factor-beta; and 8-12 parts of recombinant human insulin. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and the application thereof disclosed by the invention have the following beneficial effects: (1) the proliferation efficiency is remarkably improved; (2) the cell morphology and dryness can be well maintained; and (3) the compliance and the compatibility are good.

The application relates to a microbial fermentation device for beef jerky processing and relates to the technical field of food fermentation equipment, and comprises a material conveying pipe. The application can accurately control the temperatures of the front section, the middle section and the rear section of the material conveying pipe by arranging three groups of heating components and uniformly distributed temperature sensors, can match the growth activity peak value of probiotics, can improve the proliferation efficiency of a bacterial population and the activity of probiotics, can avoid flavor substance imbalance, and can improve the heating efficiency and uniformity by adopting an opening design of a heat preservation shell and a spiral-wound heating pipe. Meanwhile, the heat loss is reduced by a protective shell and a thermal insulation aerogel, the internal components are protected, the perforation of spiral conveying blades is matched with a turnover plate, the beef jerky material mixing is promoted, the temperature accurate control in the beef jerky fermentation process, the uniform mixing of beef jerky material and the uniform distribution of bacterial liquid are realized, the fermentation efficiency and product flavor coordination are effectively improved, the fermentation period is shortened, and the product quality is improved.

The present application relates to the fields of animal nutrition and microbial fermentation technology, and particularly relates to a strain of Saccharomyces boulardii, a microecological preparation thereof and application thereof.The Saccharomyces boulardii is named as NCUTY 002, has been preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO:M 2025648.The Saccharomyces boulardii NCUTY 002 screened in the present application has excellent acid tolerance (pH≤2.5) and bile salt tolerance (0.5%), and shows high self-aggregation capacity, so that it can survive and stably colonize in the extreme environment of digestive tract of egg ducks, and can be used as a special feed additive for egg ducks.

According to the method for inducing the illicium verum callus, the optimal induction method for rapidly growing and loose callus is screened out by exploring the optimal hormone factors and conditions for inducing the illicium verum callus, and key technical support is provided for research of an illicium verum secondary metabolite accumulation mechanism and establishment of a genetic transformation system. The invention relates to mercury bichloride, alcohol, an MS basic culture medium, 6-benzyladenine 6-BA, naphthylacetic acid NAA and zeatin ZT. ZT is one of common cytokinin for tissue culture, has strong bud induction capability, can improve the proliferation efficiency of aseptic seedlings and buds, and provides a large amount of excellent materials for callus induction.

The application discloses a kind of human mesenchymal stem cell 3D culture and amplification method, belong to stem cell culture amplification technical field.The application is prepared 3D cell culture matrix by structure design and component optimization, realize the accurate simulation of stem cell in-vivo microenvironment.The matrix is modified sodium alginate by amino hexene acid and is compounded with methacryl gelatin, and is crosslinked by lithium chloride and calcium chloride, and is synergistically formed by photocrosslinking, and the three-dimensional network has uniform pore size and excellent connectivity.The application significantly improves cell adhesion rate and proliferation efficiency, effectively maintains the biological characteristics of hMSC such as immune regulation and differentiation, the degradation rate of the matrix is dynamically matched with the cell proliferation cycle, the material has excellent biocompatibility and the degradation product is non-toxic, which solves the defects of traditional 2D culture and existing 3D technology, and provides an efficient and safe technical solution for hMSC scale-up and clinical translation.

The present application relates to stem cell technology, in particular to a pet stem cell induction medium, a preparation method and a culture method, the medium significantly improves the proliferation efficiency and multi-directional differentiation potential of adipose-derived stem cells by optimizing the ratio of specific growth factors. The preparation method comprises the steps of preparing a basic medium, adding growth factors and sterile filtering. Experimental verification shows that compared with the control group, the adipogenic differentiation efficiency of ADSCs cultured in the medium is increased by 40%, the osteogenic differentiation ALP activity is increased by 65%, and the positive rates of surface markers CD29, CD44 and CD90 are all greater than or equal to 95%. The present application solves the problems of animal source pathogen contamination risk and low induction efficiency of traditional culture medium, and is suitable for pet medical treatment.

The present application relates to a kind of preparation method for preparing neural stem cell with high proliferative capacity, belong to the field of biotechnology.The method is isolated from mammalian embryo brain tissue primary neural stem cell, with DMEM / F12 as base medium, add B27, N2, glutamine and double resistance, then join epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor-1, Y-27632 and heparin form optimized proliferation culture medium.Cell is seeded in the culture dish coated with polylysine and laminin, culture under suitable conditions, neural ball reaches certain diameter after digestion and passage.This method can improve the proliferation efficiency of neural stem cell, maintain stemness and differentiation potential, adopt serum-free system, simple operation, suitable for large-scale expansion and research application.

The present invention relates to the field of cell culture technology, and in particular to a CIK cell culture method and application thereof. The culture method comprises the following steps: (1) isolating peripheral blood mononuclear cells (PBMC); (2) preparing culture medium A; (3) preparing culture medium B; (4) resuspending and inoculating peripheral blood mononuclear cells; and (5) collecting cell culture. The culture method of the present invention can improve the yield of CIK cells, and wherein CD3 + CD56 + The proportion of double-positive cells is high, which increases cell proliferation activity and promotes the increase of CIK cell proliferation and killing activity.

This invention discloses the application of a polysaccharide coating membrane in the preparation of cell culture products, belonging to the field of cell culture food technology. Specifically, after cells are seeded onto a scaffold, a polysaccharide coating membrane is formed around it. This can alleviate the phenomenon of cells detaching from the scaffold during culture, thereby ensuring the effective proliferation and differentiation of muscle stem cells and / or adipose stem cells on the scaffold, providing a new solution for the efficient and standardized production of cell culture products.

The application discloses a tissue culture large-scale propagation method of Camellia meiocarpa. The method comprises the following steps: obtaining aseptic seedlings of Camellia meiocarpa, cluster bud proliferation culture, single plantlet culture of adventitious buds, bottle-out root preparation culture and bottle-out root culture. The composition of the cluster bud proliferation culture medium and the plantlet culture medium is optimized, the cluster bud proliferation and the formation of the adventitious buds are promoted, and the proliferation coefficient reaches 8.18. In addition, the bottle-in and bottle-out root culture scheme is combined, so that the rooting rate and the survival rate are significantly improved, and the rooting rate reaches 90%. The tissue culture method has important significance in the protection and population recovery of Camellia meiocarpa, and provides technical support for the protection and sustainable utilization of the endangered species.

The invention relates to the technical field of biological fermentation, and particularly discloses a method for preparing L-alanine through biomass fermentation. The method comprises the following steps: by taking corncobs, corns or corn straws as raw materials, sequentially carrying out acid treatment, alkali treatment and enzymolysis to obtain saccharified liquid rich in glucose, and taking the saccharified liquid as a fermentation carbon source; then carrying out seed culture on the L-alanine production strain, and carrying out immobilized packaging by adopting a sodium alginate-chitosan system to prepare an immobilized cell microcapsule; finally, the microcapsules are inoculated into a fermentation culture medium with corncob saccharification liquid as a main material for strict anaerobic fermentation, and low residual sugar concentration is maintained by feeding the saccharification liquid. According to the method, efficient saccharification and comprehensive utilization of the corn biomass are achieved, the stability and reusability of thalli in the production environment are improved through the cell immobilization technology, the production efficiency and product concentration of L-alanine are remarkably improved, and the method has the advantages of being low in raw material cost and environmentally friendly in process.

The invention relates to an efficient adventitious bud regeneration method of a new sakura variety 'Himalaya', which comprises the following steps: by taking leaves of an evergreen new variety 'Himalaya' sakura as explants, performing adventitious bud regeneration induction, cluster bud proliferation and rooting culture to obtain rooting tissue culture seedlings, and performing seedling hardening and domestication to obtain sakura seedlings. According to the method, 6-BA and NAA with a certain concentration are added into an induction culture medium, adventitious buds are induced from leaf explants through a direct organ regeneration way, the adventitious buds can form cluster buds and proliferation can be achieved by adding 6-BA, IBA and GA3 into a proliferation culture medium, the proliferation coefficient reaches about 5.5, and adventitious roots are formed at the stem bases by adding IBA and NAA into a rooting culture medium. The method is used for tissue culture and rapid propagation of a new variety of oriental cherry, the seedling propagation time is shortened, the seedling production efficiency is improved, a oriental cherry direct organ regeneration technology is established, and the oriental cherry can be used as a receptor of oriental cherry genetic transformation.

The invention provides a method for forming a regenerated plant by a direct somatic embryogenesis pathway of a pearly konjac, collecting the germination of unopened spathe flames of a pearly konjac class yellow konjac, using tender female inflorescences as explants, screening and optimizing the female inflorescences of the pearly konjac to induce direct somatic embryogenesis, maturation and plant regeneration conditions, and preliminarily establishing an efficient plant regeneration system. Under suitable temperature, light and culture conditions, the somatic embryo directly germinates into seedlings and transplants well. The invention provides a theoretical basis and technical methods for the in vitro preservation of germplasm resources of pearly konjac in Hainan hot zone, rapid breeding of seedlings and large-scale promotion, and the construction of a genetic transformation receptor system regeneration system of pearly konjac, which is of great significance to the sustainable development of the pearly konjac industry in Hainan hot zone.

The application provides a kind of festuca ovina mature seed callus induction medium, induction method and the method for establishing regeneration system, belong to plant tissue culture technical field.The application provides a kind of callus induction medium, with MS medium as base medium, further include the following components with the following content: 2, 4-dichlorophenoxyacetic acid 4~6 mg / L, hydrolyzed casein 40~60 mg / L, abscisic acid 1~3 mg / L and kinetin 0.04~0.07 mg / L.The callus induction medium described in the application can efficiently induce mature seeds to form callus, and the induction rate of callus is more than 81%.The application selects festuca ovina mature seeds as explants for callus induction culture, which is convenient and low in cost, and lays a foundation for the research of festuca ovina gene editing.