132results about How to "Improve proliferative ability" patented technology

The invention provides a novel strain M2108 of a salt-tolerant bacillus, the preservation number of the salt-tolerant bacillus is CGMCC (China General Microbiological Culture Collection Center) No.28283, and the novel strain M2108 of the salt-tolerant bacillus has relatively good salt tolerance and can grow and proliferate in a high-salt environment; the strain can be rapidly and stably colonized in a plant, and is rapid in propagation speed and strong in multiplication capacity in the plant. The new strain M2108 of the salt-tolerant bacillus provided by the invention has a treatment effect and a high control effect on various plant diseases: the control effect on diseases such as tomato root rot, damping-off and damping-off caused by fusarium oxysporum, pythium aphanidermatum and rhizoctonia solani is high, and the average control effect is 91.2% or above. When being used for preventing and treating plant diseases, the salt-tolerant bacillus M2108 is not easy to generate drug resistance, has good drug effect durability, is safe to human and livestock, and does not cause environmental pollution; the salt-tolerant bacillus M2108 microbial agent and the biocontrol agent provided by the invention are simple in preparation method, low in cost and simple and convenient to use. The salt-tolerant bacillus M2108 has a good application prospect in the prevention and control of plant diseases, and lays a foundation for the development of efficient and broad-spectrum biocontrol preparations.

The invention belongs to the technical field of stem cells, and particularly relates to a product containing mesenchymal stem cells, a preparation method, a quality control method and application in prevention and treatment of digestive system diseases. According to the preparation method, 1-3 complete culture media are provided for culturing the mesenchymal stem cells, and the obtained mesenchymal stem cells are good in subpackage form; cD73, CD90 and CD105 are highly expressed, and CD19, CD34, CD45, CD11b and HLA-DR are not expressed; the cell proliferation capacity is strong, the osteogenesis and adipogenic differentiation capacity is strong, and the medicine is safe and free of toxic and side effects. The prepared mesenchymal stem cells are high in quality, have good performance of secreting HGF, KGF and IGF-1, and can effectively treat digestive system diseases.

The invention discloses an artificial antigen presenting cell for efficiently preparing a chimeric antigen receptor T cell (CAR-T) and a construction method of the artificial antigen presenting cell, and belongs to the field of cell therapy and immunotherapy, the artificial antigen presenting cell takes a K562 cell as a platform, an anti-CD3 single-chain antibody, an anti-CD28 single-chain antibody, a membrane binding type cell factor and a costimulatory ligand are stably expressed through genetic engineering, and the chimeric antigen receptor T cell (CAR-T) is obtained. And the local concentration of the anti-CD3 antibody is improved by adopting an HOTag3 tool. The multi-signal integration strategy obviously enhances the activation and proliferation capacity of the T cells, promotes the CAR-T cells to maintain a low differentiation state, and endows the CAR-T cells with excellent anti-tumor activity and durability. The artificial antigen presenting cell provides a novel efficient tool for the field of cellular immunotherapy, and is expected to promote development of more efficient and safer CAR-T therapy to cope with challenges of various malignant tumors.

The invention discloses an induction culture medium for preparing human umbilical cord mesenchymal stem cell exosomes and application of the induction culture medium, and relates to the technical field of exosome culture. The additive is prepared from the following components: 0.8 to 1.5 percent of ITS, 0.03 to 0.05 percent of a luteolin extract, 0.04 to 0.06 percent of a herba taraxaci extract, 0.02 to 0.04 percent of a radix ginseng extract, 0.03 to 0.05 percent of resveratrol, 0.01 to 0.03 percent of vitamin C, 0.03 to 0.05 percent of a pimpinella thellungiana extract, 0.01 to 0.03 percent of a rhizoma dioscoreae extract and 0.01 to 0.02 percent of sodium citrate; the induction culture medium provided by the invention not only can avoid introduction of exogenous substances and reduce the pollution risk, but also can promote proliferation of stem cells and effectively induce synthesis and exosome secretion of a large number of human umbilical cord mesenchymal stem cells.

The invention provides a recombinant salmon mitochondrial gene (mDNA) as well as construction and application of a cloning vector of the recombinant salmon mitochondrial gene (mDNA). After the recombinant salmon mitochondrial gene (mDNA) is subjected to vector construction, cell transformation and proliferation amplification, polydeoxyribonucleotide (PDRN) with higher purity and yield can be prepared on a large scale, and compared with PDRN prepared from wild type mitochondrial DNA, the PDRN prepared from the recombinant mitochondrial gene has the advantages of oxidation resistance, inflammation resistance, improvement of cell proliferation ability, improvement of cell proliferation ability and the like. Compared with the prior art, the PDRN prepared from the salmon testis has remarkable and better effects in the aspects of resisting ultraviolet damage, perfecting the cell repair capacity and the like, the effect is equivalent to that of PDRN prepared from the traditional salmon testis, a new thought or method is provided for large-scale preparation or industrial production of polydeoxyribonucleotide, and the application prospect is wide.

The invention relates to the technical field of organoid culture, and relates to a culture system and a culture method for gastrointestinal tumor organoids for the first time, the culture system is formed by adding antioxidant active traditional Chinese medicine extracts such as naringin or polydatin on the basis of an existing culture system, when the culture system is used for culturing the gastrointestinal tumor organoid, the primary survival rate of the gastrointestinal tumor organoid and the proliferation rate after passage can be improved. Besides, the invention further provides application of the culture system of the gastrointestinal tumor organoid, the culture system and the cultured gastrointestinal tumor organoid are used for screening small-molecule targeted anti-tumor drugs, and a better in-vitro research model is provided for subsequent anti-tumor treatment.

The invention relates to the field of biological medicine, and discloses application of a biphenyl cyclooctene lignan compound and a chemotherapy sensitizing drug. The biphenyl cyclooctene lignan compound can be used for preparing a chemotherapy sensitizing drug, and the chemotherapy sensitizing drug is an ovarian cancer chemotherapy sensitizing drug, and can improve the sensitivity of cells to paclitaxel and inhibit the activity, proliferation and migration of the cells. The biphenyl cyclooctene lignan compound is combined with paclitaxel, so that the cell activity of the ovarian cancer paclitaxel drug-resistant cell A2780 / Taxol can be remarkably reduced, and the cell survival rate is only 31.13%. The combination of the biphenyl cyclooctene lignan compound and paclitaxel can also be used for preparing chemotherapy sensitizing drugs.

The invention belongs to the technical field of cell culture, and particularly relates to a culture method for in-vitro amplification of NK cells activated by NK cell exosomes, which comprises the following steps of: after promoting the NK cells to generate a large amount of exosomes by using an electrical stimulation method, filtering, performing ultrafiltration, extracting and purifying the NK cell exosomes by using specific immunomagnetic beads, thereby obtaining the NK cell exosomes activated by the NK cell exosomes. A set of strict detection and quantification standards is established for the exosome by utilizing Western Blot, a BCA method, DLS and NTA technical means, a primary PBMC is obtained by adopting a density gradient centrifugation method, and the high-activity and high-purity NK cell exosome is dynamically added into a cell culture medium in the activation and amplification stages of the PBMC, so that self-activation of the NK cells is realized, the proliferation efficiency of the NK cells is remarkably improved, and the NK cell exosome has a good application prospect. Meanwhile, the safety risk of a culture system is reduced.

The invention relates to the technical field of cell culture, and particularly provides an NK (Natural Killer) cell serum-free culture solution which is prepared from 24 percent by mass of traditional Chinese medicine extract and the balance of serum-free culture solution, the culture method of the NK cells comprises the following steps: adding the NK cells into a culture bottle, sampling and counting, diluting the cell density to 1 * 10 < 6 > / mL by using the serum-free culture solution of the NK cells, culturing in an incubator with the temperature of 37 DEG C and the CO2 content of 5% for 15 days, sampling and counting, circularly placing in a 326 mT magnetic field for 540 seconds and a 383 mT magnetic field for 120 seconds during culture, and keeping the two magnetic field environments at an interval of 0.5 hour. The serum-free culture device has a unique serum-free culture-magnetic field system, so that the multiplication capacity and killing capacity of the NK cells are not influenced or even improved while serum is not used.

The invention relates to the technical field of nano-material anti-tumor drugs, and provides a SiO2 / Ica-PDA-FA multifunctional nanoparticle and a preparation method and application thereof.The multifunctional nanoparticle is of a core-shell structure, silicon dioxide serves as a carrier, icaritin is loaded on the surface of the silicon dioxide, and the surface of the silicon dioxide is modified with polydopamine and folic acid; the problem of poor water solubility of icaritin can be improved, tumor tissues can be actively targeted through folic acid, release of icaritin is promoted in a cell acidic environment and under the condition of high-concentration glutathione, so that expression of HSP90 is inhibited, the heat-resistant characteristic of tumor cells is overcome, and the anti-tumor activity of icaritin is improved. Under the irradiation of near infrared rays, the dual anti-tumor effects of low-temperature photo-thermal treatment and icaritin drug treatment are realized.

The invention discloses a stem cell-derived function-enhanced mesenchymal stromal cell as well as a preparation method and application thereof. The preparation method comprises the following steps: inserting an IL-10 gene coding sequence and a CD47 gene coding sequence into a human pluripotent stem cell; a neural crest cell differentiation culture medium containing a Wnt pathway agonist and an ALK5 inhibitor is used for carrying out induced differentiation culture on the CD47 positive human pluripotent stem cells to obtain the neural crest cells. The neural crest cells are subjected to induced differentiation culture by adopting a transition culture medium containing an epidermal growth factor, a fibroblast growth factor 2 and a cell apoptosis inhibitor and a mesenchymal stromal cell differentiation culture medium containing the epidermal growth factor, the fibroblast growth factor 2 and a platelet growth factor in sequence; according to the present invention, with the combination of the specific embodiments, the obtained mesenchymal stromal cell highly expresses CD47 and IL-10, the expression of the CD47 can help the transplanted cell escape from the attack of the immune system and prolong the cell survival time, and the expression of the IL-10 can be used for anti-inflammation.

The invention provides a mutant spinal glioma cell line. The H3K27M mutant type glioma cell line is successfully constructed, the constructed cell line has more excellent multiplication capacity, migration capacity, tumor formation capacity and drug resistance, the curative effect and potential side effects of drugs can be more comprehensively evaluated, the cell line can be used for in-vitro research on the biological function of H3K27M glioma and response to drug treatment, and the cell line can be applied to in-vitro research of the H3K27M mutant type glioma. Wide application prospects are realized.

The invention belongs to the technical field of autoimmune skin disease immunotherapy, and provides a three-target novel chimeric autoantibody receptor structure which comprises three chimeric receptors through two connecting peptides, specifically, a first chimeric autoantibody receptor comprises a Dsg1 extracellular structural domain; the second chimeric autoantibody receptor comprises a Dsg3 extracellular structural domain; the third chimeric antigen receptor comprises an extracellular domain capable of targeting a B cell associated antigen; wherein the Dsg1 extracellular structural domain is an extracellular structural domain of desmosome core glycoprotein 1; the Dsg3 extracellular structural domain is the extracellular structural domain of desmosome core glycoprotein 3. In-vitro experiments and animal experiments verify that the novel three-target CAAR-T cells can significantly increase the release of cytokines, enhance the targeted killing activity of the cytokines, and show better multiplication capacity; a novel cellular immunotherapy technology is provided for treating autoimmune skin diseases, and the application of CAAR-T in the field of autoimmune disease treatment can be expanded.

The invention discloses a culture medium and a culture method of appendix mucinous tumor organoid. The culture medium of the appendix mucinous tumor organoid is prepared by adding the following components with final concentration into a basic culture medium: 2-8nM of NRG1 and 1-2mu g / ml of R-Spondin1, 90 to 110 [mu] g / ml of Noggin; 70 to 80 [mu] g / ml of Wnt-3a; eGF with a concentration of 45 to 55 ng / ml; the concentration of the FGF-10 is 90 to 110 ng / ml; 1X B to 27TM; 4 to 6 mM of nicotinamide; 4 to 6 nM of prostaglandin E2; 9 [mu] g / ml to 11 [mu] g / ml of Transferrin human; 4-6 [mu] g / ml of insulin; 4 to 6 nM of gastrin I; 9 to 11 [mu] M of Y27632; 1 to 2 [mu] M of A83-01; 4 to 6 [mu] M of SB202190; and 2 to 3 [mu] M of CHIR999021. The culture medium is high in induction rate.

The invention discloses a chimeric antigen receptor double-negative T cell drug co-expressing CXCR5 and IL-7 and application thereof, the cell drug can co-express CXCR5 and IL-7, and the cell drug comprises EF1-alpha, a CD8 alpha signal peptide, an NKG2D extracellular domain, a CD8 alpha hinge region, a CD8 alpha transmembrane region, a costimulatory factor 4-1BB, a CD3 zeta intracellular signal region, T2A, a chemotactic factor receptor CXCR5, P2A and interleukin IL-7 which are connected in sequence; according to the chimeric antigen receptor double-negative T cell drug co-expressing the CXCR5 and the IL-7 and the application of the chimeric antigen receptor double-negative T cell drug, the CXCR5 and the IL-7 are co-expressed on CAR-DNT cells of targeted NKG2DL, so that the migration and infiltration capacity of the drug to a tumor site is enhanced, the survival and proliferation capacity of the CAR-T cells is improved, the killing effect of the CAR-DNT cells on tumors is improved, and the drug has a good application prospect. The purpose of treating tumors is achieved, and allogenic treatment can be achieved.

The invention belongs to the field of cell culture, and discloses an osteosarcoma organ culture medium and a gelatin planting method. The following components are added into the osteosarcoma organ-like culture medium: a B27 additive, an N-2 additive, a GlutaMAX additive, casein amino acid, N-acetylcysteine, nicotinamide, vitamin C, sodium pyruvate, human gastrin, Wnt3A, R-spondin 1 protein, Noggin protein, activin A protein, Forskolin, Y-27632, SB202190, CHIR99021, A83-01, h-EGF, FGF-10, HGF, VEGF A, IGF 1, IL-6, cyan-streptomycin and fetal calf serum. The osteosarcoma organoid culture medium provided by the invention can promote organoid cells to maintain relatively high multiplication capacity and maintain self-renewal, so that the amplification efficiency and the modeling success rate of the organoid are greatly improved.

The invention belongs to the technical field of biology, and particularly relates to an immune engineering cell for expressing a discontinuous truncated functional region of GM-CSF and application. The invention firstly provides a CAR-NK cell for expressing a discontinuous truncated functional region of GM-CSF, and the CAR-NK cell expresses a novel chimeric antigen receptor structure; the novel chimeric antigen receptor structure comprises a discontinuous truncated functional domain fragment of a GM-CSF protein and a chimeric antigen receptor; the non-continuous truncated functional domain fragment of the GM-CSF protein comprises an A helix fragment, a C helix fragment and an exon 2 fragment. The invention also provides a CAR-T cell for expressing the discontinuous truncated functional region of the GM-CSF as well.

The invention relates to a mesenchymal stem cell serum-free culture medium with limited chemical components as well as application and a method thereof, the serum-free culture medium is completely serum-free and has definite components: all additives are known chemical substances and meet the production requirements of cGMP (Complementary Good Manufacturing Practice); telomere shortening can be obviously delayed; replicative senescence is slowed down by activating TERT expression; the multiplication capacity can also be enhanced; the P10 generation cells still have the active division capacity; multidirectional differentiation is supported; the osteogenic differentiation efficiency is superior to that of a traditional serum-containing system; and the method is suitable for industrial production and clinical transformation.

The invention belongs to the technical field of biology, and particularly relates to an immune engineering cell for expressing an exon 2 functional area of GM-CSF and application. The invention firstly provides a CAR-NK cell which expresses a novel chimeric antigen receptor structure; the novel chimeric antigen receptor structure comprises an exon 2 functional domain fragment of a GM-CSF protein and a chimeric antigen receptor. The invention also provides a CAR-T cell of the exon 2 functional domain fragment which also expresses the GM-CSF. The invention provides a novel chimeric antigen receptor structure modified by a fragment only containing a unique functional region in the full-length GM-CSF, which not only retains the function equivalent to that of the traditional CAR and CAR-GM, but also obtains an improved function.

The invention relates to the field of biology, in particular to a composite nutrient supplement emulsion capable of improving immunity and a preparation method thereof.According to the composite nutrient supplement emulsion capable of improving immunity, walnut peptides are further improved, the walnut peptides shown as SEQ ID NO: 8 and SEQ ID NO: 15 are screened finally, and the walnut peptides have higher DPPH and ABTS clearance rates; cell experiments prove that the walnut peptide can lower the content of IL-10 and increase the content of IL-17A, the walnut peptide can regulate inflammatory factors to achieve the purpose of improving the immunity of the organism, and mouse experiments prove that the proliferation capacity of splenic lymphocytes and the activity of NK cells of mice are remarkably improved; and a mouse experiment proves that the effect of the prepared compound nutrient supplement emulsion is better than that of the pure walnut peptide, which indicates that other components can achieve a synergistic effect with the walnut peptide. The composite nutrient supplement emulsion can improve immunity and has great economic value.

The application discloses a method for culturing three-dimensional cell spheres of alveolar type II epithelial cells under a condition without exogenous matrix glue, and comprises the following steps: A, perfusion digestion treatment is performed on mouse lung tissue to obtain a single cell suspension; B, the single cell suspension is inoculated into a culture dish coated with anti-CD31 antibody and anti-CD45 antibody for immunoadhesion culture, under the culture condition, cells expressing CD31 and CD45 are subjected to immunoadhesion, meanwhile, fibroblasts with strong adhesion ability are attached to the surface of the culture dish, so that non-epithelial cells are removed, and unattached cells are collected to obtain an enriched ATII cell population; and C, the unattached cells are inoculated into a low-adhesion culture environment for suspension culture, and the culture system contains signal regulatory factors for maintaining the stability of the ATII cell phenotype, and the enrichment degree of epithelial cells is improved, and the interference of non-target cells is reduced through the combination of negative immunoadhesion of the anti-CD31 antibody and the anti-CD45 antibody and short-time adhesion treatment.

The invention belongs to the technical field of stem cell treatment, and discloses a composition for treating degenerative arthritis and application of the composition, and the composition comprises the following components in percentage by mass: 20-60% of adipose-derived stem cells, 10-30% of bone marrow mesenchymal stem cells and 10-50% of traditional Chinese medicines. According to the application, the adipose-derived stem cells with stronger multiplication capacity are obtained by improving a culture medium of the adipose-derived stem cells, miRNA-140 of the adipose-derived stem cells is highly expressed, inflammatory factors can be effectively inhibited after the adipose-derived stem cells are compounded with bone marrow mesenchymal stem cells and turmeric powder in proportion, and the recovery of cartilage tissues is accelerated.

The present application relates to the technical field of organoid culture, and specifically discloses a culture medium for musk gland organoids and its preparation method and application. The present application discloses a culture medium, which uses DMEM / F12 as the basic culture medium; contains the following concentrations of components as functional additives: albumin, B‑27, HEPES, glutamine, N‑acetyl‑L‑cysteine, niacinamide, growth factor, Wnt3a, A83‑01, Forskolin, penicillin, streptomycin, hydrocortisone, insulin; the growth factor is EGF and bFGF with a weight ratio of 350: (11‑21). The present application also discloses a method for preparing the above-mentioned culture medium and its application in musk gland organoid culture. The culture medium of the present application can be used to obtain musk gland organoids with stem cell characteristics, and the cell activity is high.

The invention relates to a CD70 and BCMA targeting dual-targeting chimeric antigen receptor and an application thereof. The CD70 and BCMA targeting dual-targeting chimeric antigen receptor has specific binding ability to CD70 and BCMA. The immune effector cell prepared on the basis of the double-targeting chimeric antigen receptor has the following advantages that the probability that tumor cells escape treatment by losing a single antigen is greatly reduced, and the targeting treatment effect is improved; a CD70 nano antibody sequence is used, so that the length of an insertion fragment is greatly shortened, and the development difficulty of a process end is reduced; and the polypeptide has excellent proliferation capacity, is beneficial to prolonging the in-vivo duration time, and plays a more remarkable anti-tumor role. The invention provides a treatment or improvement approach for diseases related to BCMA-CD70 expression.

The application belongs to the field of treating blood vessel-bone coupling imbalance diseases in traditional Chinese medicine, and particularly relates to a composition for treating blood vessel-bone coupling imbalance diseases, a pharmaceutical preparation and application. Geniposide alone or in combination with crocin I can effectively reduce endothelial cell damage under pathological stimulation conditions, reduce the level of cell apoptosis and the degree of oxidative stress, improve the vascular endothelial function state, and can also significantly inhibit osteoclast differentiation, reduce the number of mature osteoclasts, and reduce bone resorption activity, thereby being conducive to restoring the dynamic balance between bone formation and bone resorption.

The invention discloses an NK (Natural Killer) cell in-vitro amplification culture method which comprises the following steps: on the 0th day, inoculating mononuclear cells into a culture bottle containing an NK cell culture solution, autologous plasma and an exosome of HEK293 cells, and standing and culturing in an incubator for 14 days; during the culture period, supplementing the NK cell culture solution and the autologous plasma once every two days, and controlling the cell density to be 1.0 * 10 < 6 > cells / mL; wherein the exosome of the HEK293 cell also needs to be supplemented on the seventh day; and after 14 days of culture, collecting centrifugal cells to obtain the NK cells. According to the culture method, the exosome of the HEK293 is used for replacing trophoblast cells added in the process of culturing the NK cells by a feeder method, so that the safety risk possibly caused by directly using the trophoblast cells is reduced; meanwhile, the HEK293 exosome is added and is combined with the NK cell surface receptor, so that synergistic stimulation is provided, the NK cells are activated and amplified at a high speed, and the multiplication capacity and purity of the NK cells are greatly improved.

The invention discloses a TIL cell culture medium and a TIL cell culture method, an activation and proliferation culture medium of TIL cells comprises the following components: a T cell serum-free culture medium, IL-2, platelet lysate and gentamicin, a CD3 monoclonal antibody, a CD28 monoclonal antibody and phytolectin PHA-P are added for cell activation, and then the activation and proliferation culture medium is used for activation and proliferation culture. Wherein CD3 and CD28 antibodies are used in combination to stimulate TIL cells and simulate the double-signal effect of in-vivo TIL cell activation, meanwhile, PHA-P is utilized, by means of the characteristics of PHA-P, small gonorrhea cells are stimulated in vitro to be converted into gonorrhea cells with immunocompetence, division of the gonorrhea cells is promoted, and the proliferation capacity of the TIL cells is improved through the synergistic effect of PHA-P and IL-2; platelet lysate is used for replacing plasma of a patient and is used as an efficient culture supplement for supporting TIL cell growth; finally, more TIL cells are obtained through the technology, and the number of immune cells required by clinical effective treatment can be sufficiently met.

The invention discloses an application of a high mobility group protein B2 (HMGB2) in regeneration and repair of cardiac muscle after myocardial injury, and belongs to the technical field of biomedicine. Test results show that by overexpressing HMGB2 protein in myocardial cells, the in-vitro myocardial cell proliferation capacity is remarkably enhanced, and meanwhile, after acute myocardial ischemia injury in vivo, the myocardial cells can quickly start endogenous myocardial regeneration to play roles in promoting myocardial regeneration and repair, reducing scar formation, improving cardiac functions and the like. Besides, the invention designs a drug delivery mode based on a virus vector carrying HMGB2 gene, the problems of poor targeting, low biological safety, low treatment efficiency and the like of a traditional treatment method are solved, a new drug action target is provided for prevention and treatment of acute myocardial infarction and myocardial ischemia-reperfusion injury, and the drug delivery system has important clinical transformation value.

The present invention belongs to the field of cell culture technology, and specifically relates to a method for isolating and primary culturing adult Muscovy duck cardiomyocytes. In the culturing method of the present invention, the steps for isolating and purifying adult Muscovy duck cardiomyocytes are simple, the cells adhere quickly, the survival rate is high, the activity is good, and the cell culture system has good stability. The method is a relatively ideal method for culturing primary Muscovy duck cardiomyocytes, which can basically meet the requirements of various research fields of medicine, biology, pharmacy and life sciences for experimental research related to Muscovy duck cardiomyocytes, and is suitable for promotion and application in general laboratories.

The invention relates to the technical field of biological pharmacy, in particular to application of disulfiram in preparation of medicine for treating diabetic femoral bone loss, and the femoral bone loss is caused by diabetes. It is found through experiments that disulfiram can promote the multiplication capacity of bone marrow mesenchymal stem cells damaged by diabetes and the osteogenic differentiation capacity of the bone marrow mesenchymal stem cells damaged by diabetes, inhibit the adipogenic differentiation capacity of the bone marrow mesenchymal stem cells damaged by diabetes and can promote bone mass recovery of lost femur bone caused by diabetes; the medicine can be used for treating diabetes mellitus bone mass reduction.