26 results about "Serum free medium" patented technology

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.

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 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.

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 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 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.

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 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.

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.

Disclosed herein are activators and expanders for NK cell culture, coating agents for NK cell culture vessels, serum-free media for activation and expansion steps of NK cell culture, and methods of NK cell culture.

[Problem] To provide a method for producing a culture supernatant liquid, the method making it possible to produce a large amount of a culture supernatant liquid containing an active ingredient at a high concentration. [Solution] The method for producing a culture supernatant liquid comprises: (a) a step for supplying a culture solution containing a carrier, mesenchymal stem cells, and serum (hereinafter referred to as "FBS-DMEM") to a culture tank; (b) a step for adhering the mesenchymal stem cells to the carrier; (c) a step for culturing the mesenchymal stem cells using the FBS-DMEM; (d) a step for removing the FBS-DMEM while leaving the carrier-adhered mesenchymal stem cells in a filter; (e) a step for washing the culture tank, the carrier-adhered mesenchymal stem cells, and the filter; (f) a step for supplying a serum-free culture solution for a supernatant liquid (hereinafter, referred to as "CM-DMEM") to the culture tank; (g) a step for culturing the mesenchymal stem cells using the CM-DMEM; and (h) a step for recovering the CM-DMEM while leaving the carrier-adhered mesenchymal stem cells in the filter.

The invention relates to the technical field of biological medicine, in particular to an autophagosome for promoting hair growth and an extraction method and application thereof. Comprising the following steps: inoculating human umbilical vein endothelial cells or human placenta mesenchymal cells into a DMEM (Dulbecco Modified Eagle Medium) complete medium, and culturing until the cell fusion degree reaches 80-90%; replacing the DMEM complete culture medium with a serum-free DMEM culture medium, collecting supernate after culture is completed, and centrifuging the supernate to obtain a crude extract containing the autophagosome; and resuspending the crude extract by using PBS, adding an LC3 antibody, and finely extracting by using a paramagnetic particle method to obtain the autophagosome. The method has the advantages of simple operation process, no need of other inducers in the process of inducing the formation of the autophagosome, no too high requirements on laboratories, and high yield of the autophagosome, and provides an efficient, convenient, economical and practical treatment means for alopecia patients.

The invention belongs to the technical field of poultry vaccines, and particularly relates to a chicken new tributary method adeno-quintuplet vaccine and a preparation method thereof.The chicken new tributary method adeno-quintuplet vaccine comprises chicken Newcastle disease venom, infectious bronchitis venom, avian influenza venom, infectious bursal disease virus liquid and fowl adenovirus venom, and the preparation method of the Newcastle disease venom comprises the steps that firstly, frozen MDCK suspension cells are provided; the frozen MDCK suspension cells are recovered at the temperature of 40-45 DEG C; 2, transferring the resuscitated MDCK suspension cells to a serum-free culture medium; according to the invention, the inactivated vaccine is prepared from the Newcastle disease venom, the infectious bronchitis venom, the avian influenza venom, the infectious bursal disease virus liquid and the fowl adenovirus venom by adopting a superstrong concentration process and a high-quality adjuvant, so that after animals are immunized, antibodies are generated quickly, the titer is high, the protection period is long, and the occurrence and spreading of epidemic diseases are prevented; and people can conveniently put multi-combined vaccines to cope with continuously changing epidemic disease prevention and control situations.

A synthetic meat product for human and animal consumption and methods for producing such food product are disclosed. The synthetic food product comprises or essentially consists of a cell biomass of avian cells grown in vitro in a chemically-defined serum free culture medium under controlled conditions and do not contain ant hazard contaminations.

The invention discloses a serum-free culture medium and application thereof in culture of large yellow croaker muscle stem cells. The serum-free culture medium comprises a basic culture medium and an exogenous additive component, wherein the exogenous additive components comprise HEPES, sodium chloride, hydroxypropyl-beta-cyclodextrin, sodium metavanadate, vitamin C magnesium phosphate, omega-3 polyunsaturated fatty acid, pyridoxine hydrochloride, nicotinamide, cholesterol, calcium chloride, potassium chloride, a soybean pancreatin inhibitor, taurine, trehalose, ethanolamine, a telomerase activator and recombinant human serum albumin. The animal-derived protein-free serum-free culture medium disclosed by the invention does not contain animal-derived components such as serum, does not need to add components such as growth factors or hormones, not only has definite chemical components, high safety, low cost and high cell proliferation speed, but also can effectively improve the myogenic differentiation potential of muscle stem cells and effectively activate telomerase activity to solve the problem of cell aging; and a feasible product and a feasible method are provided for culture of fish cells and large-scale production of cell culture meat.

This invention discloses a culture medium and method suitable for large-scale full-suspension culture of Vero cells. The culture medium is a serum-free medium specifically designed and developed based on the growth and metabolic characteristics of Vero suspension cells and their different proliferation abilities when cultured in a reactor versus a shaker, combined with the differences in culture scale. It includes the following components: amino acids, proteins, vitamins, inorganic salts, serum replacement factors, and auxiliary components. Furthermore, this invention determines the control parameters of the full-suspension culture reactor based on cell growth characteristics, significantly increasing the VERO cell culture density. The cell doubling rate reaches 10-fold after 48 hours of culture, the cell production cycle is halved, and production efficiency is improved. The operation is simple, reducing the contamination rate in large-scale culture and solving the problem of long perfusion culture cycles. This is of great significance for the large-scale production of viral vaccines using VERO cells as a substrate.

Provided is a serum-free formula for T cell expansion, which comprises a basal serum-free medium and a cytokine combination, and the cytokine combination comprises IL-2, IL-4, IL-7, IL-10 and IL-15; wherein, based on a total volume of the basal serum-free medium, a content of IL-2 is from 5 ng / ml to 50 ng / ml, a content of IL-4 is from 5 ng / ml to 200 ng / ml, a content of IL-7 is from 5 ng / ml to 90 ng / ml, a content of IL-10 is from 5 ng / ml to 50 ng / ml and a content of IL-15 is from 5 ng / ml to 200 ng / ml. The serum-free formula for T cell expansion can exert the effects of expanding T cells, especially selectively expanding the CD3+CD8+ T cells, under a serum-free circumstance, thereby avoiding the uncertainty and the risks derived from the animal serum and promoting the development of adoptive cell therapy or immunotherapy.

The invention relates to the technical field of cell culture, in particular to a culture medium and a culture method of gamma delta T cells. In order to solve the problems of large batch-to-batch production difference, high biosafety risk, unstable supply chain and high cost caused by combined use of serum or blood substitution and multiple cytokines in the existing gamma delta T cell culture process, the serum-free culture medium containing GlutaMAX and IL-2 is obtained through screening, and the culture medium is applied to gamma delta T cell culture and has the advantages that the culture medium has a wide application prospect. The method can improve the process batch stability, can realize linear amplification, has higher purity and amplification efficiency, reduces the biological safety risk, can significantly reduce the supervision and production cost, and meets the GMP consistency requirement.

The present invention aims to provide a method that makes it possible to sufficiently proliferate cells, particularly immunocompetent cells, even in a serum-free medium or low-serum medium, and a medium additive and a medium composition to be used for the method. An effect of maintaining and promoting cell proliferation comparable to that achieved by culture in a serum-containing medium can be obtained even in a serum-free medium or a low-serum medium, by containing boric acid or a salt thereof.

Provided are an activator and an expanding agent for NK cell culture, a coating agent for an NK cell culture container, a serum-free medium for an activation step and an expansion step of NK cell culture, and an NK cell culture method.

The application discloses a serum-free culture medium for vascular endothelial cells and application thereof, and belongs to the technical field of cell culture. The serum-free culture medium for vascular endothelial cells comprises a basic culture medium, and further comprises human serum albumin, endothelial cell growth factor, epidermal growth factor, follicular inhibin-like recombinant protein 1, fibroblast growth factor, insulin-like growth factor, heparin sodium, hydrocortisone, L-glutamine and ascorbic acid. The serum-free culture medium has the advantages of clear components, stable quality and low cost, can promote the division and proliferation of vascular endothelial cells, can better maintain the state of endothelial cells, has high scientific research and medical application values, and can make the cultured vascular endothelial cells have the advantages of high proliferation speed, short doubling time, good cell state, high survival rate and high activity. In addition, the serum-free culture medium does not contain xenoantigen components, will not cause an immune rejection reaction with the human body, and is suitable for industrialized production of vascular endothelial cell related products.