55 results about "Serum free medium" patented technology

The invention relates to a culture medium and a culture method of persistent DC-CIK cells and application of the culture medium and the culture method, and belongs to the technical field of cell culture. The invention provides a culture medium for persistent DC-CIK cells, which takes a serum-free culture medium as a solvent and comprises a recombinant human granulocyte macrophage colony stimulating factor, a recombinant human interleukin-12, a recombinant human tumor necrosis factor-alpha, a recombinant human interferon-gamma, L-cysteine, glutathione, ginseng polysaccharide and streptomycin sulfate. According to the culture medium and the culture method of the persistent DC-CIK cells, proliferation and growth of the DC-CIK cells can be promoted, and very high tumor killing activity of the DC-CIK cells can be maintained for a long time.

The invention discloses a cord blood-derived regulatory T cell amplification culture medium and a use method thereof, and relates to the field of animal cell culture and immune cell therapy, the culture medium is a serum-free culture medium, the culture medium is composed of a basic culture medium, a serum substitute combination, a combination of four cell factors, an immunosuppressor, an apparent stabilizer and an antioxidant, and all the components play a role synergistically; according to the use method, cell amplification is realized through a collaborative process of staged environmental regulation and targeted activation. According to the method, the problems of exogenous pollution and batch difference caused by dependence of a serum-containing system on cord blood-derived regulatory T cell amplification in the prior art are solved, the defect that a serum-free scheme is difficult to consider the amplification efficiency and the function stability at the same time is overcome, the cell amplification quality and the clinical application safety are guaranteed, and the method is suitable for large-scale clinical transformation requirements.

A method for domesticating CHO-K1 cells into high-expression cells, comprising the following steps: culturing adherent CHO-K1 cells in a complete culture medium, to obtain log growth CHO-K1 cells; transferring the log growth CHO-K1 cells to a serum-free medium for suspension culture; performing a first liquid replacement domestication, to obtain domesticated CHO-K1 cells; placing the domesticated CHO-K1 cells in a chemically defined medium for suspension culture; performing a second liquid replacement domestication, to obtain domesticated suspended CHO-K1 cells; performing monoclonalization on the domesticated suspended CHO-K1 cells, to obtain CHO-K1 monoclonal cells; and screening the CHO-K1 monoclonal cells, to obtain high-expression CHO-K1 monoclonal cells. This method can domesticate CHO-K1 original parent cells into suspended cells, and screen cells having higher expression capability from the domesticated cell population, thereby improving expression levels.

The invention discloses a ferroptosis inhibitor and application of the ferroptosis inhibitor in radiation pneumonitis drugs, and relates to the technical field of biological medicines. The specific preparation method comprises the following steps: adding the ferroptosis inhibitor and the auxiliary agent into the solvent, and dissolving to obtain the ferroptosis inhibitor composition; wherein the ferroptosis inhibitor is isoorientin; the auxiliary agent is a glucosamine derivative; the solvent is a serum-free DMEM culture medium. The ferroptosis inhibitor composition prepared by the method provided by the invention can effectively reduce the rise of active oxygen, ferrous ions and malondialdehyde levels of human pulmonary epithelial cells BEAS-2b and mouse pulmonary epithelial cells MLE-12 caused by irradiation, restore cell mitochondrial membrane potential enrichment and inhibit the expression of various inflammatory factors in irradiated cells; therefore, the process of radiation pneumonitis is remarkably improved, and a new medication strategy is provided for research on radiation pneumonitis drugs.

The present invention provides a method for producing a retinal progenitor cell, including (1) a first step of subjecting pluripotent stem cells to floating culture in a serum-free medium to form an aggregate of pluripotent stem cells, and (2) a second step of subjecting the aggregate formed in step (1) to floating culture in a serum-free medium or serum-containing medium each being free of a substance acting on the Sonic hedgehog signal transduction pathway but containing a substance acting on the BMP signal transduction pathway, thereby obtaining an aggregate containing retinal progenitor cells.

The invention relates to a Vero cell serum-free culture medium and application thereof, the serum-free culture medium is suitable for free suspension growth of Vero cells and is free of animal sources and protein components, adherent cells can be directly domesticated through a certain method to screen out Vero suspension cells, and serum-free suspension culture of the Vero suspension cells is achieved in the serum-free culture medium. The density of cells subjected to suspension culture by adopting the serum-free culture medium can reach more than 107 cells / mL, the cell viability is more than 90%, the cells are well dispersed, the cells can still normally grow and proliferate after continuous subculture for 20 generations, and the cells are sensitive to various viruses.

The present invention provides: a stable fibrin clot-soluble protein excellent in production in protein-expressing cells in a serum-free medium and suppressing the production of impurities such as degradation products; a pharmaceutical formulation containing the protein; and a production method for the protein and pharmaceutical formulation.

The invention discloses an umbilical cord mesenchymal stem cell serum-free culture medium, which is composed of a basic culture medium and additive components, and the additive components comprise, by final concentration, 5-20 [mu] g / mL of astragalus polysaccharide, 0.5-2.0 [mu] g / mL of salidroside, 0.1-0.5 [mu] g / mL of tanshinone IIA, 0.1-0.3 mg / mL of sodium pyruvate, 5-15 mM of D-ribose, 3-6 mg / mL of recombinant human albumin, and 5-10 [mu] g / mL of recombinant transferrin; the invention also discloses a method for culturing umbilical cord mesenchymal stem cells under the hypoxia condition by using the serum-free culture medium. The method comprises the steps of cell inoculation, culture under the hypoxia condition, subculture, cell passage and the like. The astragalus polysaccharide, the salidroside, the tanshinone IIA, the sodium pyruvate, the recombinant human albumin and the recombinant transferrin are added into the serum-free culture medium, so that nutrition supply and function maintenance of cells under a low-oxygen condition are guaranteed, the proliferation rate and the differentiation capacity of the cultured cells are improved, the apoptosis rate of the cells is reduced, and the survival rate of the cells is increased. Wide application prospects are realized in the fields of cell culture, regenerative medicine and the like.

Provided is a serum-free medium that is composed of a fewer number of constituent components and is less expensive.　The serum-free medium comprises Dulbecco's modified Eagle's medium (DMEM), albumin (BSA), and insulin-transferrin-selenium-ethanolamine (ITS-X). The serum-free medium is suitable for the culturing of mouse ES cells that are naïve pluripotent stem cells, and bears comparison with the most superior medium on the market.

The invention discloses an NK cell culture solution and a culture method thereof. The culture solution comprises an amplification culture solution and an induction activation culture solution, wherein the amplification culture solution is based on a serum-free culture medium and is added with cell factors such as IL-2, IL-12, IL-15 and IL-18, and the induction activation culture solution is based on the amplification culture solution and is added with induction activation factors such as OK432, CD16-alpha, a pseudomonas aeruginosa injection, alpha-galactosylceramide and CD137L. The induction activation factor CD16-alpha added in the NK cell culture fluid can activate NK cells independently or in combination with OK432, a pseudomonas aeruginosa injection, alpha-galactosylceramide and the like, so that the NK cells grow quickly, and the CD3-C56 + ratio of the NK cells can be increased; after the CD16-alpha is combined with the CD137L, the activated NK cells can be further activated, so that the activity of the NK cells is higher, and the killing effect is stronger.

The present invention provides a method for producing a retinal progenitor cell, including (1) a first step of subjecting pluripotent stem cells to floating culture in a serum-free medium to form an aggregate of pluripotent stem cells, and (2) a second step of subjecting the aggregate formed in step (1) to floating culture in a serum-free medium or serum-containing medium each being free of a substance acting on the Sonic hedgehog signal transduction pathway but containing a substance acting on the BMP signal transduction pathway, thereby obtaining an aggregate containing retinal progenitor cells.

The invention relates to the technical field of biological medicine, and provides application of a dental pulp stem cell exosome in preparation of a medicine for treating and preventing osteoarthritis, a preparation method of the dental pulp stem cell exosome comprises the following steps: S1, obtaining dental pulp tissue from a dental pulp cavity of an in-vitro tooth; s2, pretreating dental pulp tissues, adding digestive juice, digesting, centrifuging, discarding supernatant, adding a complete culture medium containing antibiotics into precipitates, mixing, filtering and centrifuging to obtain dental pulp stem cells; and S3, culturing the dental pulp stem cells by using a complete culture medium, when the confluence degree of the dental pulp stem cells reaches 60-80%, replacing the serum-free culture medium for continuous culture, collecting the dental pulp stem cell conditioned culture medium, and performing differential centrifugation to obtain the dental pulp stem cell exosome. According to the technical scheme, the regulating effect of the dental pulp stem cell exosome on endoplasmic reticulum stress and the mechanism of the dental pulp stem cell exosome for improving cartilage cell degeneration are explored.

A lentiviral vector production system comprises (a) a lentiviral culture supplement to control cell growth, (b) a transfection reagent comprising DHDMS, DOPE, and cholesterol to increase transfection efficiency, (c) a lentiviral production enhancer comprising sodium propionate, sodium butyrate, and caffeine to boost lentiviral production, wherein the lentiviral vector production system is serum-free. A method of lentiviral vector production comprises using the lentiviral production system. Another method for lentiviral vector production comprises (a) culturing eukaryotic cells in a serum-free medium, (b) providing a lentiviral culture supplement to control cell growth, (c) transfecting the cells with a lentiviral vector using a transfection reagent comprising DHDMS, DOPE, and cholesterol to increase transfection efficiency, and (d) providing a lentiviral production using a lentiviral production enhancer comprising sodium propionate, sodium butyrate capable of boosting lentiviral production.

To provide methods for producing an artificial skeletal muscle tissue from pluripotent stem cells.SOLUTION: The present invention describes a method for preparing an engineered skeletal muscle tissue, as well as skeletal myoblasts, skeletal myotubes, and satellite cells, a medium used being serum-free and different chemicals and their concentrations, as well as the physical stimuli, being defined. Additionally, the methods described herein are performed without transfection of human cells with transgenes. The engineered skeletal muscle cells exhibit myoblast-, myotube-, or satellite cell-specific genetic markers, demonstrating efficient differentiation of these cell types. Despite its engineered production, the skeletal muscle tissue possesses excellent stimulus-dependent contractility and exhibits contractions in response to different stimulation frequencies.SELECTED DRAWING: None

This invention proposes a method for preparing dental pulp stem cell exosomes and their applications, belonging to the field of exosome technology. Dental pulp stem cells from exfoliated deciduous teeth are seeded into serum-free α-MEM medium containing microparticles, insulin-like growth factor, transforming growth factor-β1, penicillin, and streptomycin for culture. Exosomes are isolated, dispersed in PBS solution, and incubated with microparticles, propylene glycol, and rhodioloside to obtain highly active dental pulp stem cell exosomes. The surface of these exosomes is then modified with shikonin and interferon to obtain dental pulp stem cell exosomes. The dental pulp stem cell exosomes prepared by this invention exhibit good affinity for titanium implant-mediated osteointegration repair materials, demonstrating good anti-inflammatory, immunomodulatory, osteogenic, and damage-reducing effects. Furthermore, the preparation method is simple, yields high output, and operates under mild conditions, showing broad application prospects.

The invention provides a dental pulp stem cell exosome as well as a preparation method and application thereof, and belongs to the technical field of exosomes. The method comprises the following steps: inoculating exfoliated deciduous tooth pulp stem cells into a serum-free alpha-MEM culture medium containing microparticles, insulin-like growth factors, transforming growth factor-beta1, penicillin and streptomycin for culturing, separating exosomes, dispersing the exosomes into a PBS solution, adding microparticles, propylene glycol and salidroside for incubation, and separating to obtain the high-activity pulp stem cell exosomes. And performing surface modification on alkannic acid and interferon to prepare the dental pulp stem cell exosome. The dental pulp stem cell exosome prepared by the invention has relatively good affinity to a titanium implant material mediated osseointegration repair material, has relatively good advantages of resisting inflammation, regulating immunity, promoting osteogenesis, reducing injury and the like, is simple in preparation method, relatively high in yield and mild in condition, and has a wide application prospect.

The invention discloses an application of acetic acid-2-(1H-indole-3-yl)-ethyl ester in preparation of antitumor drugs, and belongs to the technical field of chemical combination drugs, SiHa, HCC94, CaSki human cervical squamous carcinoma cells, Hela human cervical adenocarcinoma cells, SKOV3 human ovarian cancer cells and Hep-G2 human hepatoma cells are inoculated into a 96-well plate, 5000 cells exist in each well, and the cells are inoculated into the 96-well plate. On the second day, replacing the culture medium with a complete culture medium containing compounds with different concentrations, incubating in the incubator for 48 hours, and replacing the culture medium with a serum-free culture medium containing 10% CCK8 solution; according to the present invention, the light absorption value of the microplate reader at the wavelength of 450 nm is determined after the incubation in the incubator for 1 h, and the compound has effects of inhibition on the proliferation of a variety of tumor cells, such that the compound acetic acid-2-(1H-indole-3-yl)-ethyl ester can be used for preparing the cancer treatment drug, and has good development and application prospects.

The method of hair transplant using dermal papilla and dermal sheath obtained from hair follicle as disclosed comprises the steps of: providing initial keratinocytes to be used for creating condition medium; creating and collecting condition medium from said keratinocytes; collecting hair follicle cells containing dermal papilla and dermal sheath from patients; culturing said cells as obtained from patients using condition medium collected from the prior step; injecting cultured cells in the predetermined amount and the predetermined ratio of dermal papilla cells and dermal sheath cells in the predetermined amount area characterized in that the step of injecting and collecting condition medium from said keratinocytes comprises the step of applying a solution obtained from the mixture of defined keratinocyte-serum free medium (DK-SFM) and Mesenchymal stem cell culture medium (MSC) to said initial keratinocytes, removing defined keratinocyte-serum free medium (DK-SFM), and extracting a condition medium from the said mixture and wherein the step of culturing hair follicle cells as obtained from patients comprises the step of applying condition medium extracted and MSC in the ratio of 1:1.

The application belongs to the technical field of biological medicine, and particularly relates to umbilical cord mesenchymal stem cells and a preparation method and application thereof. The preparation method takes umbilical cord Wharton's jelly as raw material, and obtains high-purity umbilical cord mesenchymal stem cells through cleaning treatment, tissue block preparation, serum-free culture and liquid change subculture. The culture conditions and operation parameters are strictly controlled in the preparation process, the MSC serum-free culture medium and trypsin substitutes are used, so that the cell activity and purity can be guaranteed. The umbilical cord mesenchymal stem cells have no obvious toxicity and tumorigenic risk under a safe dose, and have no damage to organs in long-term use. The umbilical cord mesenchymal stem cells can significantly improve the motor dysfunction after cerebral ischemia, reduce the volume of cerebral ischemic injury, protect the integrity of the blood-brain barrier, reduce the neuroinflammatory response, and play a multi-dimensional role in brain tissue protection and repair, thereby providing a new effective means for the treatment of ischemic stroke, and having important clinical transformation value and application prospect.

The present invention proposes a highly active product obtained based on antler stem cells, a preparation method and its application, including preparing a serum-free culture medium, thawing the immortalized antler stem cells, centrifuging, inoculating and culturing, and passage, and finally resuspending and freeze-drying with water for injection to obtain freeze-dried powder. The culture medium consists of a basic culture medium and a growth additive, and each component is described in detail. The method can efficiently enrich the active products unique to antler wax flakes. It also involves the application of highly active products obtained based on antler stem cells in skin care products (such as water solutions, lotions, etc.) and health products (such as nutritional supplements, functional drinks), providing a new way for the development of related products, and has potential economic and social value.

The invention belongs to the technical field of biology, and discloses a preparation method and application of a stem cell preparation for repairing ovarian function premature senility, the preparation method comprises the following steps: adding stem cells and vascular endothelial growth factors into a serum-free medium for culturing to obtain a proliferated stem cell solution; the amino component, the aldehyde component and the connexin are added into normal saline according to a preset proportion to be mixed, hydrogel is obtained, and an imine group formed by amino in the amino component and aldehyde in the aldehyde component is broken under the preset stress condition; and stirring and mixing the hydrogel and the proliferated stem cell solution to obtain the stem cell preparation. By means of the preparation method, the homing rate of the stem cells in the injection process for treating the premature ovarian failure is increased, and the loss rate of the stem cells is reduced.

According to the method for differentiating the SH-SY5Y cells into the neuron-like cells, rapid proliferation of the cells is facilitated in a DMEM culture medium containing 10% of serum, a differentiation reagent EC23 is used when differentiation begins, a DMEM / F12 culture medium with the concentration of 5% of serum is replaced, cell differentiation is facilitated, and finally, a serum-free culture medium NBA containing a differentiation reagent BDNF is used for differentiating the cells into the neuron-like cells. Compared with the prior art, the neuron-like cells with high SH-SY5Y differentiation efficiency can be rapidly obtained, a network is formed between the improved neuron-like cells, the survival rate of the cells is also greatly improved, and the neuron-like cells can be better used for research of subsequent experiments.

Provided are a serum-free medium for co-incubation of tumor organoids and immune cells, a tumor organoid immune microenvironment system and a construction method therefor, and a method for evaluating the efficacy of drugs based on tumor organoid immune co-culturing. The used medium for co-incubation of tumor organoids and immune cells can well maintain the growth of tumor organoids and immune cells, and can provide nutritional support for these two different types of cells. In addition, the established method for evaluating the efficacy of drugs in a tumor organoid immune co-culture system can be used for evaluating the efficacy and toxic side effects of an immune drug against a tumor, helping to select potentially effective drugs and optimize a treatment scheme. By means of the organoid immune co-culture system, the response of different patients to drugs can be better understood, providing a basis for personalized medicine and precision treatment.

The invention relates to the technical field of biological medicine, in particular to a serum-free culture medium suitable for menstrual blood stem cell amplification and a preparation method and application thereof.The serum-free culture medium is prepared from, by mass, 90-110 parts of a basic culture medium, 1-1.6 parts of a serum substitute, 0.28-0.34 part of a metabolism regulator, 0.00008-0.00012 part of an antioxidant, 5-20 ng / mL of growth factors FGF-2 and 1-2 ng / mL of TGF-beta1. The preparation method comprises the steps of mixing the components, filtering and sterilizing, and aseptically adding the growth factors. According to the culture medium, through the synergistic effect of key components, the proliferation efficiency of menstrual blood stem cells is remarkably improved, apoptosis is inhibited, cell dryness is maintained, meanwhile, the serum pollution risk is thoroughly eliminated, and the culture medium is suitable for large-scale production and cell therapy application.

To provide a serum-free formulation for T cell expansion.SOLUTION: A basal serum-free medium and a combination of cytokines, wherein the combination of cytokines comprises IL-2, IL-4, IL-7, IL-10, and IL-15, wherein, based on the total volume of the basal serum-free medium: A serum free formulation for T-cell expansion is provided wherein the amount of IL-2 is between 5ng / ml and 50ng / ml, the amount of IL-4 is between 5ng / ml and 200ng / ml, the amount of IL-7 is between 5ng / ml and 90ng / ml, the amount of IL-10 is between 5ng / ml and 50ng / ml, and the amount of IL-15 is between 5ng / ml and / ml. 200ng.EFFECT: The serum-free formulation for T-cell proliferation exerts an effect of proliferating T-cells under serum-free conditions, especially selectively proliferating CD3 + CD8 + T-cells, thereby avoiding uncertainties and risks derived from animal serum and facilitating the development of adoptive cell therapy or immunotherapy.SELECTED DRAWING: None

The invention relates to the technical field of mesenchymal stem cell culture, in particular to a mesenchymal stem cell culture medium and a culture method. The mesenchymal stem cell culture medium is a serum-free culture medium formed by taking a DMEM / F12 culture medium as a basic culture medium and adding supplemental factors, the components in the culture medium are clear, adverse factors such as batch-to-batch quality difference and serum-derived pollution caused by a traditional serum-containing culture medium can be effectively avoided, and the culture medium has the advantages that the culture medium is suitable for large-scale popularization and application. Therefore, the influence of serum components on experimental research is avoided, and a positive effect on ensuring the accuracy of experimental research is achieved; meanwhile, corn glycopeptide is added into the culture medium, on one hand, a nitrogen source needed by cell proliferation is supplemented, on the other hand, rich amino acid and glucose can be provided, and the positive and effective effects on stable and rapid growth and proliferation of cells are achieved.

The invention relates to neurobiology, molecular virology, biomedical engineering and the like, and discloses a domesticated low-serum Vero cell line and a method for preparing an HSV virus vector through suspension culture cell amplification. Aiming at the bottleneck problem that a process for amplifying HSV by adherent culture of Vero cells is difficult to realize large-scale and engineering amplified production, the invention researches and develops a method for preparing HSV based on suspension culture of cell amplification. Vero cells introduced from ATCC are domesticated, and the serum content of a culture medium is gradually reduced, so that the Vero cells are suitable for suspension culture in a microcarrier or sheet carrier reactor by using a low-serum or serum-free culture medium. The method comprises the following steps: using Tamp of Dibide Bioengineering (Shanghai) Co., Ltd; and carrying out suspension culture of the Vero cells and infection amplification of the HSV virus vector by using a J Cloud Ready TM bioreactor. High-activity suspension culture and HSV amplification of Vero cells are realized by optimizing production conditions, cell density, biochemical detection and the like, and a stable and efficient method is provided for preparing a high-titer and high-purity HSV virus vector in combination with purification process upgrading.

This application relates to the field of cell preservation, and discloses a long-term preservation medium for stem cells based on a three-dimensional biomimetic microenvironment, which comprises the following component materials in parts by mass: flexible skeleton, benzoyl peroxide, superoxide dismutase, retinoic acid, ferulic acid, mixed buffer, nucleic acid aptamer targeting stem cells, serum-free medium, bFGF and TGF-β. Its construction method comprises the following steps: S1, three-dimensional scaffold construction; S2, antioxidant loading; S3, buffer preparation; S4, aptamer modification; S5, medium integration. The three-dimensional scaffold structure constructed by natural polysaccharides and nanocomposites, combined with the biomimetic pore and flexible network design, effectively simulates the extracellular matrix microenvironment. At the same time, by improving the structural stability and chemical compatibility of the cell adhesion interface, the cell morphology maintenance and adhesion force maintenance during the freezing process are realized, so that the scaffold provides a dynamically adaptable stable environment for cells structurally.

The application discloses a kind of serum-free medium for efficiently enriching tumor stem cells and application thereof, belong to the technical field of cell biology.The serum-free medium includes basic medium, basic additive factor, functional additive component;Among them, functional additive component includes: 1 μM-1mM of alpha-ketoglutaric acid dimethyl ester, 10nM-10 μM of butyl benzyl phthalate, 0.01 μM-1 μM of endothelin-1, 0.01mM-1mM of uridine diphosphate glucose, 1nM-100nM of tetrabromobisphenol A.The application provides stable stemness maintenance conditions for tumor stem cells by adding functional additive component to serum-free medium;At the same time, the synergistic effect between functional additive component and other components significantly improves the enrichment efficiency of tumor stem cells, and has the advantage of low cost;Therefore, it has good application prospect in efficient enrichment of tumor stem cells.

The invention discloses a stem cell preparation as well as a preparation method and application thereof in anti-aging repair, and relates to the field of biological medicines. The stem cell preparation comprises the following components: a 3D mesenchymal stem cell suspension, a probiotic-plant fermentation supernatant, a targeted delivery medium and a cryopreservation protection matrix. The preparation method comprises the following steps: inoculating mesenchymal stem cells into a bioreactor in a serum-free culture medium, and carrying out suspension culture to obtain a 3D mesenchymal stem cell suspension; preparing probiotic-plant fermentation supernate on a basic culture medium through double-bacterium fermentation; and after mixing, adding a cryopreservation protection matrix, and after programmed cooling, performing liquid nitrogen cryopreservation. The stem cell preparation can be applied to preparation of anti-aging repair drugs. The stem cell preparation disclosed by the invention can solve the retention problem, break through concentration contradiction, overcome the burst release defect and reverse recovery injury; the traditional Chinese medicine composition has obvious effects on repairing senescence skin and improving senescence cardiac hypofunction.