323 results about "Basal medium" patented technology

The invention discloses a sheep formation state pluripotent stem cell culture medium and application thereof. The invention belongs to the technical field of biology, and particularly relates to a sheep formation state pluripotent stem cell culture medium and application thereof. The composition for culturing the sheep formation state pluripotent stem cells, disclosed by the invention, contains an N2B27 basic culture medium and cell factors, wherein the cell factors contain CHIR99021, IWR1, A419259, a recovery hub IL6, a recovery hub sIL6R, a recovery hub Activin A and a recovery hub FGF2 (Fibroblast Growth Factor 2). The culture medium prepared from the composition can obtain stable sfPSCs, and can be applied to construction of sheep gene editing model animals.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a device and a method for gradually adjusting the glucose concentration of an in-vitro embryo culture solution. The device comprises at least two miniature liquid storage bins, wherein each miniature liquid storage bin is filled with a high-concentration glucose solution in advance; the slow release channel is respectively connected with each micro liquid storage bin and the culture solution main container; the culture solution main container is used for accommodating an embryo and a basal culture medium; the control unit is used for sequentially opening the release ports of the miniature liquid storage bins, so that the high-concentration glucose solution sequentially flows into the culture solution main container through the slow release channels. The technical means of staged closed-loop micro sugar control is introduced into the structure and the using method of the device, so that the accurate adjustment of the sugar concentration of the in-vitro embryo culture solution is realized, the requirements of embryos in different development stages can be effectively met, and the embryo culture quality and the subsequent development potential are remarkably improved.

The invention relates to the technical field of cell culture. The invention provides a mesenchymal stem cell culture medium without animal-derived components and application of the mesenchymal stem cell culture medium. The culture medium comprises a basic culture medium and an additive, the additives comprise FGF-2, IGF-1, glutamine, folic acid, serotonin, cholesterol, arachidonic acid, an HEPES buffer solution, sodium bicarbonate, SCF, TGF-beta, PDGF, ascorbic acid and transferrin. The mesenchymal stem cell culture medium disclosed by the invention is completely free of animal-derived components, and meets the requirements of FDA / EMA clinical-grade cell therapy products. The culture medium can support high-density amplification of mesenchymal stem cells, the activity of passage cells is effectively improved, and the amplification time is shortened. The method has remarkable significance in improving the amplification efficiency of the mesenchymal stem cells and promoting and applying factory growth.

The invention relates to an N2B27 culture medium for inducing generation of bidirectional pluripotent stem cells, one of LY2090314, AS1842856 or CHIR99021 is added into an N2B27 basal culture medium, and the concentration of the CHIR99021 is 10 mu M. The invention also relates to a preparation method of the N2B27 culture medium. The weight of the LY2090314 is 10 nM, and the weight of the AS1842856 is 0.6 [mu] M. The invention also provides an application of the culture medium in induced production of bidirectional pluripotent stem cells. According to the invention, a novel stem cell culture condition for promoting co-expression of genes OCT4 and CDX2 by adding small molecules LY2090314 (LY), AS1842856 (AS) and CHIR99021 to inhibit signal channels of FOXO1 and GSK3 is screened out. The culture conditions are simple and convenient, the application range is wide, the cultured cells keep the characteristics of 16-32 cell stages of the embryos, and the development characteristics of the embryos can be efficiently reproduced. Under the condition of not depending on transcription factors, the screened bidirectional pluripotent stem cells (BPSCs) can promote efficient generation of TSC cells and establish a TSC cell line through independent differentiation of a serum culture medium or induced differentiation of a TSC culture medium. The obstacles between early embryo pedigree are broken, and the research on early embryo development is promoted.

Techniques for generating and utilizing adult hepatocyte organoids are described. An example method includes generating an organoid by culturing hepatocytes obtained from an adult donor in a hepatocyte culture medium. The hepatocyte culture medium includes a basal medium and 5 μM of a TGF-β inhibitor.

The invention discloses a culture medium composition for culturing GDT cells and a culture method, and belongs to the field of biological medicine. Aiming at the problems of low in-vitro amplification efficiency and poor function of the existing GDT cells, the invention provides the culture medium composition for culturing the GDT cells, the culture medium composition comprises an induction culture medium and a proliferation culture medium, the induction culture medium comprises a basic culture medium and an activating agent, and the activating agent comprises IL-7 and paclitaxel; the proliferation culture medium comprises a basic culture medium, IL-18 and a metabolism regulator. An STAT5 pathway is activated through IL-7, and amplification of the Vdelta2 subgroup is specifically promoted; on the other hand, through the cooperation of the IL-7 and the IL-18, the IL-18 activates the expression of the NKG2D, and the IL-18 and the IL-7 jointly enhance the cytotoxicity; efficient amplification and function enhancement of the GDT cells are achieved; the killing rate of PD-L1 + tumor cells is larger than 80%, the motility rate after cryopreservation recovery is larger than 90%, the function maintenance rate is larger than 85%, and the cell line is suitable for tumor immunotherapy, infectious disease treatment and cell drug development.

The invention provides an osteogenic induced differentiation kit, a culture medium and a preparation method of the culture medium, and particularly relates to the technical field of stem cells. The kit comprises a DMEM (dulbecco's modified eagle medium) basal culture medium, fetal calf serum, dexamethasone, vitamin C, beta-glycerol phosphate, mesenchymal stem cell osteogenesis induced supernatant, alizarin red S staining solution and acetic acid. According to the kit provided by the invention, the problems of slow calcium nodule formation, small calcium nodule quantity and small calcium nodule in the osteogenic induced differentiation of the umbilical cord mesenchymal stem cells in the prior art are solved, so that the calcium nodule formation of the umbilical cord mesenchymal stem cells is effectively promoted, meanwhile, the calcium nodule quantity can be increased, and the osteogenic induced differentiation period is shortened.

The invention discloses a culture system, culture method and application of gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system comprises: a basic culture medium, which is a DMEM / F12 culture medium, and is additionally added with a special growth promoting factor obtained from a natural extract; the multiple growth factors comprise an epidermal growth factor (EGF), a fibroblast growth factor (FGF), a bone morphogenetic protein (BMP) and a vascular endothelial growth factor receptor inhibitor (VEGFRi), and the concentration of each growth factor is accurately screened and optimized; the extracellular matrix is Matrigel modified by physical and chemical methods; the specific additive comprises an antioxidant N-acetylcysteine (NAC). According to the culture system, the culture method and the application of the gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system which is closer to a real environment in vivo is constructed by optimizing basic culture medium components, setting various growth factors and specific additive concentrations and adopting modified Matrigel as an extracellular matrix.

The invention belongs to the technical field of biomedicine, and relates to a method for increasing the yield of pancreas islet organs differentiated by pluripotent stem cells and improving the stability. The method comprises the following steps: (1) inoculating pluripotent stem cells, and culturing by adopting a basic culture medium containing Y-27632; (2) changing the liquid, and continuously culturing by adopting a basic culture medium; (3) culturing for 1 day by using an S1D1 culture medium, and then culturing for 2-3 days by using an S1D2 culture medium; (4) changing the liquid, and culturing for 2-3 days by adopting the S2 culture medium; (5) changing the liquid, and culturing for 1-2 days by adopting an S3 culture medium; (6) changing the liquid, and culturing for 4-7 days by adopting a culture medium S4; (7) changing the liquid, and culturing for 3-10 days by adopting an S5 culture medium; and (8) carrying out cell sphere repolymerization, and culturing for 3-21 days by using an S6 culture medium to obtain the pancreas islet organ. The present invention increases the yield and differentiation efficiency at each differentiation stage and increases the scale yield at the cellular level.

Platelet lysate compositions and cell culture media compositions for maintaining and / or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and / or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and / or flasks pretreated with the cell culture media compositions.

The present invention discloses a low - serum medium and its application in the preparation of cultured meat from Larimichthys crocea cells, which is also applicable to other marine or freshwater fish, belonging to the field of bioengineering technology. The low - serum medium comprises a basal medium, fetal bovine serum accounting for 30 - 80 μL / mL, and exogenous additives. The exogenous additives include human serum albumin, ascorbic acid, basic fibroblast growth factor polypeptide, human holo - transferrin, lysophosphatidic acid, and human recombinant IGF - 1. Based on the DMEM / F12 medium, the low - serum medium provided by the present invention adds six substances that promote cell growth, reduces the dependence of satellite cells on fetal bovine serum, effectively reduces the usage amount of fetal bovine serum, and saves the culture cost. Compared with the general - type medium (DMEM high - glucose, adding 10% FBS), the low - serum medium provided by the present invention has an equivalent effect on the maintenance and proliferation ability of cell stem cells.

The present invention relates to the field of cell culture, and particularly to a serum-free induction adipogenic differentiation medium for fish stem cells and its application. It is composed of a basal medium and a combination of cell culture cofactors; the basal medium is one of DMEM and DMEM / F12 medium; the combination of cell culture cofactors consists of oleic acid, soybean lecithin, vitamin C phosphate, β-cyclodextrin, transferrin, saikosaponin A, bakuchiol, dehydroabietic acid, and phenyl lactic acid. By adding the combination of cell culture cofactors, the present invention replaces the serum in the traditional adipogenic induction differentiation medium for preadipocytes and the core component dexamethasone IBMX in the traditional adipogenic differentiation "cocktail", and obtains a medium suitable for serum-free induction adipogenic differentiation of fish stem cells, which not only solves the bottleneck problems of the prior art, but also provides reliable technical support for the large-scale production of cell-cultured fish meat.

The invention provides a culture medium composition and a method for preparing mesenchymal stem cells. The culture medium combination comprises a culture medium A, a culture medium B and a differential culture medium, the culture medium A comprises a basic culture medium and a first small molecule compound combination, the culture medium B comprises a basic culture medium and a second small molecule compound combination, and the differential culture medium comprises a basic culture medium and a third small molecule compound combination. According to the culture medium and the method, the MSC can be prepared through ES or iPSC, and the culture medium and the method can be widely applied to the fields of regenerative medicine, tissue engineering, immunotherapy and the like.

The invention discloses an application of a composition of L-arginine and L-histidine in improving the yield of N-acetylglucosamine produced by microbial fermentation. The invention also discloses a culture medium for fermentation production of N-acetylglucosamine. The culture medium comprises a basic culture medium and a fed-batch culture medium, wherein the fed culture medium is prepared from the following components: 600 to 800 g / L of glucose, 4 to 6 g / L of monopotassium phosphate, 80 to 120 g / L of glycerol, 0.8 to 1.5 g / L of L-arginine and 0.8 to 1.5 g / L of L-histidine. According to the method, L-arginine and L-histidine with specific concentrations are added in the microbial fermentation process for producing N-acetylglucosamine, so that the yield of N-acetylglucosamine can be remarkably increased.

The invention relates to a culture medium for separating and culturing stem cells in tissues and application of the culture medium. The culture medium is a basic culture medium containing genipin, plasmalogen and cell factors. The brand-new culture medium is designed, genipin, plasmalogen and cell factors are added into the basic culture medium and are integrally and synergistically matched, stem cells in tissues (such as nasal mucosa tissues) can be quickly activated, the adherence and migration speed of the stem cells is remarkably increased, proliferation of the stem cells is promoted, and cell activity is effectively maintained; the activation and separation efficiency of the stem cells is obviously improved.

The invention relates to an NK cell culture medium and an NK cell culture method, and relates to the technical field of cell culture.The NK cell culture medium comprises a basic culture medium and an additive, and the additive is prepared from, by concentration, 200-400 IU / L of cytokines, 3-6 mmol / L of nutritional supplements, 1-2 mmol / L of metabolism regulators, 90-110 mg / L of vitamin C, 150-300 micrograms / mL of plant polysaccharides, 1-2 micrograms / mL of 1, 2-propylene glycol, 1-2 micrograms / L of 1, 3-propylene glycol and 1, 3-propylene glycol. The medicine is prepared from 50-100 [mu] mol / L of 1, 3-dicaffeoylquinic acid, 48-52 [mu] g / mL of nano-scale hydroxyapatite, 14-16 [mu] mol / L of a signal channel regulator, 5-10 IU / L of heparin sodium and 10-80 [mu] g / mL of astragaloside IV-liposome. The culture medium and the culture method can efficiently amplify the NK cells and maintain the activity and the functional stability of the NK cells.

The invention relates to the technical field of cell biology, in particular to a culture medium for inducing neural stem cells and application thereof.The culture medium for inducing the neural stem cells comprises a neural ectoderm induction culture medium and a neural stem cell induction culture medium; the neural ectoderm induction culture medium is prepared from a basic culture medium, an N-2 additive, a B-27 additive, an inhibitor SB431542 and an inhibitor Dorsomorphin, the neural stem cell induction culture medium is prepared from a basic culture medium, an N-2 additive and a B-27 additive, the basic culture medium is composed of a DMEM / F12 culture medium, a Neurobase culture medium, GlutaMax and non-essential amino acid NEAA. The problems that neural stem cells obtained through an existing single-layer adherent induction method based on small molecules or growth factors are uneven in differentiation and low in purity are solved.

The invention discloses a culture medium for constructing a colorectal cancer peritoneal metastasis micro-tumor cell model and application of the culture medium. The invention relates to the technical field of biology, and provides a culture medium for culturing a micro-tumor model for colorectal cancer peritoneal metastasis. The invention discloses a cholera toxin immunopotentiator, which is composed of an antibacterial and antifungal agent three antibody, HEPES, GlutaMax, a human recombinant protein EGF, a human recombinant protein bFGF, a human recombinant protein HGF, a human recombinant protein Noggin, a human recombinant protein R-spondin 1, a human recombinant protein GDNF, SB202190, A83-01, Primocin, N-acetyl-L-cysteine, nicotinamide, an N2 additive, cholera toxin, B27, ITS-X, Y-27632, Galunissertib and a basal culture medium. The culture medium and the culture method disclosed by the invention can effectively improve the culture success rate of colorectal cancer peritoneal metastasis and focus combination.

The invention belongs to the field of stem cell culture, and relates to a pluripotent stem cell culture medium, which comprises an E6 basal culture medium, 2 ng / mL of TGF-beta1, 0-50 ng / mL of FGF-2, and 0.01-1 [mu] M of SUN11602. According to the technical scheme, proliferation and pluripotency of the hPSC are effectively maintained in a mode of combining small chemical molecules with low-concentration FGF-2.

The invention provides a culture method and an acclimatization and transplantation method for coptis chinensis test-tube plantlets, and belongs to the technical field of plant tissue culture. The method comprises the following steps: (1) disinfecting a coptis chinensis explant, inoculating the disinfected coptis chinensis explant into an MS basal culture medium, and culturing to obtain a sterile seedling; (2) inoculating the aseptic seedlings into an adventitious bud culture medium for adventitious bud culture to obtain adventitious buds; (3) inoculating the adventitious buds into a proliferation culture medium for proliferation culture to obtain proliferation seedlings; and (4) inoculating the proliferation seedlings into a rooting culture medium for rooting culture to obtain the coptis chinensis test-tube plantlets. When the adventitious bud, proliferation and rooting culture medium is used for culturing the coptis chinensis test-tube plantlet, the proliferation coefficient of the test-tube plantlet reaches 3.0, the rooting rate reaches 96.3%, and the survival rate of acclimatization and transplantation reaches 91.5% through seedling acclimatization and transplantation. The culture method provided by the invention is used for preserving the germplasm resources of the coptis chinensis, and provides a basic guarantee for the germplasm improvement of the subsequent medicinal material coptis chinensis.

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 are a novel high-concentration cell preservation liquid and a high-concentration cell preservation method.　The cell preservation liquid is for preserving cells in a non-frozen state. The cell preservation liquid contains a basal medium and an additive added to the basal medium. The basal medium is a culture medium used for cell culture. The additive contains at least genetically modified albumin and ascorbic acid (ascorbic acid derivative). The cell preservation liquid stores the cells at a cell concentration of at least 1 × 107 cells / mL. For example, when cells are preserved in a cell preservation liquid at 4°C at a cell concentration of 1 × 108 cells / mL, the cell survival rate when 72 hours has elapsed is 80% or more.

A method for producing mycelium biomass. The method comprises obtaining a base media composition and an antifoam solution; sterilizing a mixture of the base media composition and the antifoam solution

A method for high-density fermentation production of ULP involves culturing yeast containing a ULP expression vector in a basal medium comprising a carbon source (glycerol) and a nitrogen source (histidine and / or ammonium dihydrogen phosphate). This invention, through high-density yeast fermentation and employing an improved culture medium and process, significantly improves the yield and quality of ULP. The final expression level is 3-5 times higher than that achieved by conventional E. coli fermentation methods, reducing ULP production costs by at least 25%. This reduces the cost of related recombinant protein drugs and allows for wider application of this high-quality system in recombinant protein drug development.

The application belongs to the field of biological medicine, and discloses a culture medium, which comprises a basic culture medium, and the basic culture medium is added with ITS-A: 1x, arginine: 15-20 mu g / ml, SPP1: 3-5 mu g / ml and S100A9: 3-5 mu g / ml. The culture medium is particularly suitable for microtumor culture of MTC model and has good cell viability. Meanwhile, the application also provides application of the culture medium and a microtumor culture method.

The invention provides a preparation method of efficient activated NK cells, and belongs to the technical field of NK cell culture, and the preparation method comprises the following steps: S1, centrifuging whole blood, removing upper plasma to obtain a cell part, and diluting with a reagent I to obtain a cell diluent; s2, adding the cell diluent obtained in the step S1 into a centrifugal tube filled with a lymphocyte separating medium, centrifuging, sucking a middle mononuclear cell layer, and cleaning with a reagent II for multiple times to obtain target cells; s3, inoculating the target cells obtained in the S2 into a cell culture dish or a cell culture bottle, adding a lymphocyte basal culture medium and IL-2, activating the NK cells according to a traditional culture method, adding stem cell source exosomes on the seventh day of activated culture of the NK cells, adding the stem cell source exosomes once every three days later, continuously culturing to the 12th-17th day, and collecting to obtain the activated NK cells. The tumor killing effect is enhanced, and meanwhile the immune function of NK cells is adjusted.

This invention discloses a NK cell culture kit, its culture method, and its application. The kit includes an amplification medium, a high-efficiency induction medium, an NK-A coating solution, an NK-B mixture, and an NK-C mixture. The amplification medium contains basal medium, NAD+, human serum albumin, transferrin, β-glucan, glutathione, β-mercaptoethanol, and linoleic acid. The high-efficiency induction medium, in addition to the amplification medium components, contains soybean peptides, nicotinamide, inulin, and N-acetyl-L-cysteine. The NK-A coating solution contains heparin sodium, monoclonal antibodies, and antibodies. The NK-B mixture contains Inbakicept, IL-2, and IL-15. The NK-C mixture contains linolenic acid, IL-2, and IL-18. The Inbakicept factor in the kit can activate NK cells and enhance their cytotoxicity; linolenic acid can inhibit T cell proliferation, increase NK cell purity, and enhance cell efficacy.

The invention relates to the technical field of biology, and discloses a construction method of a gastric cancer organoid culture system, which comprises the following steps: preparation of a conditioned medium: culturing L-WRN cells and collecting the conditioned medium; preparing a gastric cancer organoid culture medium, and mixing the conditioned culture medium, the basic culture medium, a plurality of growth factors and additives; and extraction and culture of organoid: separating cells from gastric cancer tissues, mixing the cells with matrigel, and culturing by using the prepared culture medium. According to the method, the repeatability and the stability of the organoid model are improved, the response consistency of the organoid in drug screening is improved, and the reliability of the model in disease mechanism research and personalized medical application is guaranteed.

The application discloses a kind of preparation and suspension culture of blackland callus method, belong to the field of industrial biotechnology.It includes the following steps: after sterilization, take flower stem, access pH value is 5-7 in induction medium, under the condition of temperature is 20-30 DEG C and no light, induction culture 15-21 days, namely obtain blackland callus, wherein, the induction medium is made of MS basic medium, sucrose 25-35g / L, agar 6-7g / L, 6-BA 1.0-2mg / L, NAA 0.5-1.5mg / L, 2, 4-D 0.3-1.0mg / L and coconut water 0.5-1.5g / L composition.Under aseptic conditions, clamp the growth state of good blackland embryogenic callus 30g, inoculation is carried out in 200mL / 500mL pH value is 6.0 in MS liquid medium, temperature 25 DEG C, 110 rpm full light oscillation culture, subculture period 7 days, when subculture, the mixture of cell and culture fluid is mixed with fresh culture fluid according to 1:1 volume ratio, after equal division, continue to culture, namely obtain blackland suspension cell, wherein, the liquid medium is made of MS basic medium, sucrose 30g / L, 6-BA 1.5mg / L, NAA 0.5mg / L, 2, 4-D 0.5mg / L and coconut water 1.5g / L composition.The callus out of the rate of 88.6% is realized, and the induction rate of embryogenic callus that can be suspended culture is 68.6%-91.8%, and good callus state is provided for large-scale culture blackland cell.

The invention discloses an improved culture medium containing egg yolk protein and a chicken embryo extract and a preparation method of the improved culture medium. The culture medium is mainly composed of egg yolk protein, a chicken embryo extract and a basic culture medium component according to a specific proportion. The improved culture medium is rich in various nutritional ingredients such as proteins, amino acids, vitamins and cell growth factors, can effectively promote the growth and proliferation of various cell lines and improve the efficiency and quality of cell culture, can be widely applied to the field of in-vitro culture of animal cells, provides a choice with higher quality and better cost effectiveness for cell culture, and has a wide application prospect. The problems that a traditional culture medium is insufficient in nutritional ingredients, poor in cell culture effect and the like are solved.