1313 results about "Basal medium" patented technology

Methods and materials for culturing primate-derived primordial stem cells are described. In one embodiment, a cell culture medium for growing primate-derived primordial stem cells in a substantially undifferentiated state is provided which includes a low osmotic pressure, low endotoxin basic medium that is effective to support the growth of primate-derived primordial stem cells. The basic medium is combined with a nutrient serum effective to support the growth of primate-derived primordial stem cells and a substrate selected from the group consisting of feeder cells and an extracellular matrix component derived from feeder cells. The medium further includes non-essential amino acids, an anti-oxidant, and a first growth factor selected from the group consisting of nucleosides and a pyruvate salt.

The invention belongs to the technical fields of environmental engineering and bioengineering and particularly relates to a sphingomonas strain and applications thereof in the aspects of shortcut nitrification-denitrification of nitrogen-containing industrial waste water and domestic sewage and treatment of a polluted water source. The sphingomonas strain is separated from a Taihu water in China,is a local strain and has high safety; and the strain can be grown in a basic medium, wherein in the basic medium, CO2 is used as a carbon source and energy or CO2 and an organism are used as a mixedcarbon source and energy, and ammonia nitrogen or nitrate nitrogen is used as a nitrogen source. The bacterium liquid, the dormancy cell and the immobilized strain of the sphingomonas can decompose ammonia nitrogen into nitrite nitrogen and simultaneously can decompose the ammonia nitrogen into nitrogen, can be used as denitrification microorganism for converting the ammonia nitrogen into the nitrite nitrogen and the nitrogen, thereby achieving the shortcut nitrification-denitrification. The strain can rapidly decompose the ammonia nitrogen and is suitable for treating the nitrogen-containingindustrial waste water and domestic sewage as well as polluted water source.

The present invention belongs to the field of biotechnology, and discloses a serum-free culture medium with specific chemical compositions for in vitro culture and amplification of bone marrow mesenchymal stem cells. By adding insulin, transferrin, ethanolamine, sodium selenite, growth factors, adherent factors, hormone, putrescine, inorganic salt, vitamin, albumin and antioxidant into a basic culture medium, the bone marrow mesenchymal stem cells can attach to the culture medium under a serum free condition, so the in vitro culture and amplification are realized, the potential of multi-directional differentiation is maintained, and the amplified cells can be induced to be osteoblast and lipocyte in vitro. The serum-free culture medium has the advantages that the clinic level cell products for human produced by the serum free culture medium can effectively avoid the potential risk of producing cell products by serum culture medium. The drawing appended is a photo of the confluence of the bone marrow mesenchymal stem cells cultured by the serum-free culture medium.

The invention relates to a human mesenchymal stem cell culture medium. According to the culture medium, the basal culture medium comprises the following components based on the final concentration: 1-2g / L of human serum albumin, 5-10mg / L of transferring, 2-8mg / L of fibronectin, 1-4mg / L of laminin, 50g / L of Fe(NO3)3.9H2O, 417g / L of FeSO4.7H2O, 1-3mu g / L of estradiol, 2-5mu g / L of testosterone, 1-3mu g / L of progesterone, 39.25-117.74 mu g / L of dexamethasone, 5-10mg / L of insulin, 376.36mg / L of riboflavin, 80.96-242.87mg / L of coenzyme A, 4.41-6.17mg / L of butanediamine, 1-2mg / L of taurine, 0.61-1.85mg / L of aminoethanol, 8.81-26.42mg / L of pyruvic acid, 3.78-7.56mu g / L of sodium selenate, 292.3-584.6mg / L of L-glutamine, 2-8mu g / L of vascular endothelial growth factor, 4-10mu g / L of epidermal growth factor, 4-10mu g / L of basic fibroblast growth factor, 1-5mu g / L of leukaemia inhibitory factor, 1-5mu g / L of insulin-like growth factor-I and 2-8mu g / L of stem cell factor. The culture medium does not contain the animal serum, the potential animal endogenous endotoxin or virus of the animal serum is eliminated, and the culture medium is conveniently applied to clinics.

To provide an aqueous solution for cell preservation which is free of a natural animal-derived component such as a basal medium or serum. An aqueous preservation solution showing a high cell survival rate was obtained by removing a natural animal-derived component such as a basal medium or serum and controlling other components and their concentrations.

Fully defined media that support pluripotent cell viability, proliferation, cloning, and derivation, as well as methods and compositions including these media are described. Methods for deriving iPS cells from adult individuals under defined, xeno-free conditions are also described.

The invention relates to a culture medium for preparing neural stem cells and an application of the culture medium. The culture medium for preparing the neural stem cells comprises a culture medium suitable for the growth of stem cells, and a cell signal pathway inhibitor which is selected from at least one of the following agents: a GSK (Glaxo Smith Klein) inhibitor, an MEK (Methyl Ethyl Ketone) inhibitor, a TGF (Transforming Growth Factor)-beta inhibitor, a ROCK inhibitor and a BMP inhibitor. The somatic cells are cultured by using the culture medium, especially the somatic cells for expressing the transcription regulator are cultured, so as to effectively transdifferentiate the somatic cells into neutral stem cells and the transdifferentiation time is greatly shortened.

The invention relates to the field of cell and tissue culture, in particular to a culture medium for culturing mesenchymal stem cells. Aiming at the problems that the mesenchymal stem cells exist in an existing bovine serum culture medium and a human serum culture medium, the invention provides the culture medium for culturing the mesenchymal stem cells. According to the technical scheme, the culture medium for culturing the mesenchymal stem cells provided by the invention is characterized by comprising two kinds of components, namely a basal culture medium and culture medium additives; the additives comprise a cell growth factor combination, insulin, trace elements, lipid materials, vitamin substances, hormone-like substances, protein molecules, amino acid substances and other compounds. According to the culture medium for culturing the mesenchymal stem cells, provided by the invention, the zoonosis and the risk of human immune responses in the cell therapy process can be avoided, and the possibility that the natures of the cultured mesenchymal stem cells are different due to the difference among culture system batches can be avoided; the culture medium is suitable for culturing all the mesenchymal stem cells and is broad in prospect.

The invention provides a serum-free medium for umbilical cord mesenchymal stem cells and belongs to the technical field of cell culture. The serum-free medium for umbilical cord mesenchymal stem cells comprises a basal culture medium body and added ingredients, wherein the basal culture medium body is a DMEM culture medium; the added ingredients include a basic fibroblast growth factor hFGF, a epidermal growth factor hEGF, insulin hI, a leukaemia inhibitory factor hLIF and astragalus polysaccharide. The umbilical cord mesenchymal stem cells are subjected to subculture and amplification in the medium, and the surfaces of the cultured cells are marked and analyzed. The serum-free medium for umbilical cord mesenchymal stem cells overcomes the defect of exogenous pollution of serum and solves the problem of contradiction between the cell expansion number and cost reduction. The medium is free of serum, thereby preventing influence of animal-derived serum ingredients on cell culture. The medium can be used for studying the differentiation and proliferation adjusting mechanism of the umbilical cord mesenchymal stem cells.

The invention discloses a cell cryopreservation fluid which can be used for performing cryopreservation on mesenchymal stem cells or other cells. The cell cryopreservation fluid is prepared from the following ingredients: PBS or normal saline or a basal culture medium serving as a main ingredient, as well as one or more of polyethylene glycol, propanediol, Ectoin, albumin, trehalose, proline and poloxamer 188 which serve as additive ingredients. The cell cryopreservation fluid disclosed by the invention does not contain serums and DMSO, have the definite additive ingredients, and is controllable in quality, high in batch stability and high in safety. By virtue of preserving cells with the cell cryopreservation fluid, revived cells are high in survival rate, and the cellular morphology, the multiplication capacity and the differentiation potential are not influenced; the cell cryopreservation fluid can be used for replacing cryopreservation fluids containing the serums and the DMSO.

Compositions and methods for isolating and expanding human mesenchymal stem / progenitor cells through multiple passages in defined serum-free environments are provided. The culture media compositions includes a basal medium supplemented with a nutrient mixture such as Ham's F12 nutrient mixture, glutamine, buffer solutions such as sodium bicarbonate and hepes, serum albumin, a lipid mixture, insulin, transferrin, putrescine, progesterone, fetuin, hydrocortisone, ascorbic acid or its analogues such as ascorbic acid-2-phosphate, fibroblast growth factor and transforming growth factor β, and are free of serum or other undefined serum substitutes such as platelet lysate. Methods employing these compositions and protein-coated surfaces for the isolation of mesenchymal stem / progenitor cells from human bone marrow and other tissues such as adipose tissue are also provided. Finally, methods are also provided for serially expanding these cells through multiple passages without losing mesenchymal stem cell-specific proliferative, phenotypical and differentiation characteristics.

The invention provides a mycoplasma hyopneumoniae culture medium and a preparation method thereof, belonging to the technical field of veterinary biology. The mycoplasma hyopneumoniae liquid culture medium comprises the components as follows: brain heart infusion, lactalbumin hydrolysate, PPLO (pleuropneumonia-like organism) broth, yeast extract powder, proteose peptone, sodium thiosulfate, Hank's liquid, sodium pyruvate, 0.1% phenol red solution, penicillin and deionized water. The preparation method comprises the following steps of: adding health horse serum before using, and adding agar into the liquid culture medium to obtain a solid culture medium of mycoplasma hyopneumoniae. The viable bacteria titer of the mycoplasma hyopneumoniae culture medium can reach 1*109CCU / ml-1*1010CCU / ml; the viable bacteria titer and the separation sensibility are far higher than those of the existing culture medium, and the mycoplasma hyopneumoniae is fast in growth speed and high in the separation sensibility; and the preparation method of the culture medium is simple in technology, strong in operability, and suitable for industrial large-scale production.

The invention discloses a quantitative determination method for activity of a nerve growth factor. The method comprises the following steps of: (1) washing TF-1 cells in a logarithmic phase by a basal culture medium that contains no serum and recombined human granulocyte-macrophage colony stimulating factors, and then resuspending the TF-1 cells to obtain TF-1cell suspension liquid; (2) respectively adding the TF-1cell suspension liquid into a gradient-diluted nerve growth factor standard sample and a sample to be tested, and incubating at 37 DEG C in presence of 5% of CO2; then respectively adding an indicating agent to detect the absorbance / fluorescence value and drawing a standard curve line; and (3) calculating the activity concentration of the nerve growth factor of the sample to be tested according to the standard curve line and the absorbance / fluorescence value of the sample. The quantitative determination method for activity of the nerve growth factor disclosed by the invention has good accuracy and precision.

The invention relates to a use of a mesenchymal stem cell conditioned culture medium for cosmetics, preferably a use for skin repair and regeneration, whitening, wrinkle resisting and oxidation resisting, and is characterized in that the mesenchymal stem cell conditioned culture medium is prepared by the following method: (1) obtaining mesenchymal stem cells; (2) culturing the mesenchymal stem cells obtained in the step (1) in a complete culture medium; (3) culturing the mesenchymal stem cells obtained in the step (2) in a base culture medium; and (4) centrifuging to remove the mesenchymal stem cells, taking a supernatant, and filtering the supernatant, to obtain the filtrate which is the mesenchymal stem cell conditioned culture medium.

The present invention relates to a low serum efficiency mycoplasma gallisepticam attenuated strain culture medium and a preparation method thereof, and belongs to the technical field of veterinary biology. The culture medium comprises: (1) a base culture medium; and (2) an auxiliary culture medium, wherein the auxiliary culture medium mainly comprises MEM, yeast extract powder, tryptone, glucose, an inorganic salt, and the like, and growth, high titer and stability of the semi-finished product can be ensured with the auxiliary culture medium. According to the present invention, a titer of the semi-finished product bacterial liquid prepared by using the preparation method is up to 10<11> CCU / ml; and the culture medium adopts reduced pig serum to culture mycoplasma gallisepticam so as to reduce allergic stress reactions on chicken by heterologous pig serum, consider animal biosafety, improve antigen titer, and reduce production cost.

The invention relates to an immune cell cryopreservation solution and application thereof. The immune cell cryopreservation solution is prepared from 250-200 U / mL of recombinant human interleukin-2, 0.1-0.4 g / mL of polyethylene glycol, 0.1-0.4 g / mL 1,2-propylene glycol and 90-99% basal culture medium or sodium chloride for injection by volume. By using the immune cell cryopreservation solution for immune cell cryopreservation, the survival rate of recovery cells can reach 93% or above, the biological property of the cells is not changed, and the biological activity of the immune cells is ensured; in addition, due to the fact that no animal serum is contained in the immune cell cryopreservation solution, foreign protein cannot be introduced, the possibility of animal pathogeny contamination is reduced, and the problems that immune cells cannot be stored and transported for a long time are effectively solved.

The technological process includes feeding concentrated energy source matter fluid and amino acid, and perfusing basic culture medium. The process parameter control based on the growth and metabolism requirement of specific cell includes setting upper and lower limit indexes of glutamine, glucose, ammonia and lactic acid, controlling the feeding rate of glutamine, glucose, amino acid and other nutritive matter, and perfusing serum-free basic culture medium to lower the density of metabolic product. The cell growth state is monitored via detecting oxygen intaking rate, and the feeding rate is controlled precisely with computer program controlled feeding system. The technology of the present invention is especially suitable for large scale culture of hybrid tumor cell for producing treating antibody, and has the features o high stability, high control ability, determined control indexes, etc.

The invention discloses a stem cell medium and an application thereof. The medium comprises a basic medium, fetal bovine serum, cytokine or protein polypeptide, vitamin and lipid. The medium provided by the invention can quickly amplify stem cells and does not influence the potential of the stem cells, and the amplification velocity of the stem cells is improved by 3-5 times than a conventional medium; and moreover, the medium can be used for culturing the stem cells of various tissues and has excellent applicability, the cultured stem cells have strong differentiation capability and can be differentiated into cells with various functions; therefore, the stem cell medium has high scientific and research and medical application values.

The invention discloses a serum-free complete medium for mesenchymal stem cell, consisting of a basal medium and an additive component, wherein the basal medium is DMEM / F12 (dulbecco modified eagle medium), and the additive component consists of recombinant human insulin, human serum albumin, human transferrin, cholesterol, sodium selenite, ferric nitrate and glucan. The complete medium is free from serum, so that the defects that the serum-containing medium is unstable in serum batches, high in cell toxicity, abundant in heterologous protein and the like can be greatly avoided, and a good tool can be provided for the basic research and the clinic treatment of the mesenchymal stem cell; and the invention further achieves the effect which is approximate or equal to the serum-containing complete medium and the other serum-free complete mediums in the aspect of cell proliferation, so that the long-term subcultring of the cell can be supported.

The invention provides a culture system which contains defined components and efficiently acquires induced pluripotent stem cells (iPS cells) from somatic cells. The culture medium additive provided by the invention contains vitamin C, vitamin B12, insulin, glycogen synthase kinase-3 inhibitor, receptor tyrosine kinase and antioxidant. The culture medium additive provided by the invention can also contain a substituted serum cell growth promoter. The invention also provides a complete medium acquired by inducing pluripotent stem cells, which is prepared from one or more of basic culture medium, serum and substituted serum additive and the culture medium additive. The culture system provided by the invention does not have serum, has no animal source pollution, contains defined chemical components, efficiently acquires iPS cells, can maintain cell growth and proliferation in the process of conversion from somatic cells to iPS cells under the condition that feeder cells do not exist, simultaneously accelerates the induction course of the iPS cells obviously, and greatly improves the efficiency of inducing the somatic cells into the iPS cells.

The invention opened non-serum mediums which can proper to culture many kinds of the animal cells. The character of it is to use the DMEM / F12 as the base medium, then add the growth factor, hormone, the essential amino acid and the microelement. It has the functions: (1) it can support many cells and clones; (2) it is same as the serum medium in the growth of the cell and the expression of the product; (3) it support the long heritable culture; (4) it is benefit for the isolation of the product because it contains less protein; (5) it is cheap. We can get the high density of the cells and the product concentration using the medium.

The invention discloses an inducing culture medium for inducing fibroblast to trans-differentiate into cardiac muscle cells, a method and an application of the inducing culture medium. The inducing culture medium comprises a basic culture medium and an inducing small molecular assembly which is 6TCFOW or SCFOV, wherein 6 is E61541, T is tranylcypromine, C is CHIR99021, F is forskolin, O is Dorsomorphin, W is IWR-I, S is SB431542, and V is valproic acid. The inducing culture medium can trans-differentiate the fibroblast into the cardiac muscle cells which have normal cardiac muscle cell specific molecular tags and a normal cardiac muscle function, so that a new way is provided for solving the cell source problem of the regenerative medicine.

The invention relates to a method for rapidly cultivating chlorella and belongs to the technical field of biology. The cultivation method comprises the following steps: preparing a standard BG-11 culture medium to serve as a basal culture medium of chlorella by taking fresh running water as a solvent; adding a certain amount of organic carbon source and NaHCO3 into the standard culture medium, so that the final concentration is respectively 0.5-1.0g / L and 0.2-0.5g / L; putting the culture medium into a culture vessel, inoculating 10-20 percent of chlorella solution, sealing the opening, and culturing the chlorella solution in an environment with the illumination intensity of 3000-4000Lx, the light-dark ratio of 12h:12h and the temperature of 22-28 DEG C. According to the method, the cell density of the chlorella can reach a high level in a short cultivating cycle and is increased by about six times compared with that cultivated by using the basal culture medium. Meanwhile, the method has the advantages that the process is easy to operate, the culture medium is not required to be sterilized, the pH value of the culture medium is not required to be regulated, and the method can be widely applied to cultivating chlorella in industrial production.

The invention discloses a high-density culture method for bacilli and a culture medium. The high-density culture method for bacilli comprises the following steps including Step 1, strain domestication, Step 2, shake flask seed culture, Step 3, seed tank seed culture, and Step 4, fermentation tank high-density fermentation culture. Ingredients of a fermentation culture basic culture medium comprise peptone, yeast powder soaking, glucose, disodium hydrogen phosphate, monopotassium phosphate, magnesium sulfate, manganese sulfate, calcium chloride and bean pulp. A fermentation culture medium supplemental culture medium is glucose solution, the glucose content in the glucose solution is 3 to 4 percent of the total quantity of the fermentation liquid, and the high-density culture method for bacilli is applicable to the high-density culture of bacillus subtilis and bacillus licheniformis. The high-density culture method for bacilli provided by the invention has the advantages that the spore forming rate of the bacilli in the fermentation liquid can be higher than 90 percent, and the spore density is higher than 120 hundred million / milliliter to the lowest degree.

The invention discloses a method for optimizing a potato isolated culture one-step seedling culture medium. The culture medium makes leaves of a potato test tube plantlet undergo isolated culture to form a seedling in one step. The one-step seedling culture medium takes an MS culture medium as a basal medium, and 6-benzyladenine, naphthyl acetic acid and 2,4-dichlorphenoxyacetic acid with different concentrations and combinations are supplied in 1L of the MS culture medium; adventitious buds and adventitious roots are directly differentiated through callus induction; and the potato regenerated seedling is obtained in one step. The callus obtained by induction in a primary culture medium is unnecessarily inoculated to a differential medium for regenerating the seedling. Compared with an isolated culture multi-step regenerated seedling method, the method of the invention has the advantages of simplified steps, short culture period, high repeatability, and high seedling survival rate.

The invention discloses a modified medium for improving propagation of stems of Dendrobium officinale and a propagation method. The medium comprises MS (Murashige and Skoog) basal medium, cane sugar 20g / l, agar 6g / L, 6-BA (benayl aminopurine) 2mg / l, NAA (alpha-naphthaleneacetic acid) 0.2-0.5mg / l, banana mash 15-20g / l, potato mash 15-20g / l, and common tap water replacing distilled water. The modified MS medium is used to propagate the stems with axillary buds of Dendrobium officinale, alkaloid and polysaccharide content of Dendrobium officinale is evidently increased, and Dendrobium officinale grow normally. By the method, consumption of cane sugar is reduced, the tap water is used to replace the distilled water, and accordingly tissue culture technology for Dendrobium officinale is more convenient, more efficient and resource-saving.

The invention discloses a culture solution for inducing mesenchymal stem cells to differentiate into islet-like cells, and an inducing method and application of the culture solution. The culture solution comprises the following materials: niacinamide, Conophylline, cell growth factor, betacellulin and a base medium. The base medium contains 97% of high glucose DMEM (Dulbecco Modified Eagle Medium), 2% of B-27 and 1% of N-2. The inducing method comprises the following steps of: preparing an inducing culture solution; preparing the human mesenchymal stem cells; taking the human mesenchymal stem cells, inoculating the human mesenchymal stem cells into a six-hole ultralow absorption culture plate by 1.5-2*105cells / hole, adding 3ml inducing culture medium into each hole and carrying out suspended induction; and changing liquid at every 3 days, collecting cell supernatant at the ninth day, and storing the cell supernatant at the temperature of -20 DEG C. The culture solution disclosed by the invention has the advantages that the human mesenchymal stem cells are induced to differentiate into the islet-like cells by utilizing the combination of the niacinamide and the Conophylline, so that the inducing cycle is shortened, the suspension cells are beneficial to being clustered to form cell clusters similar to natural islets, further the induced differentiation efficiency is obviously increased and the clinical application risk is reduced; and the function of inducing the secretion of the cell insulin is obviously improved.

The present invention relates to methods and media for single cell serum free culture of CHO cells. Generally, the invention relates to a method of culturing CHO cells at a cell density of less than 100 cells / ml in a serum-free cell culture medium. The medium is sufficient to support the growth of a single CHO cell and comprises a basal medium sufficient to support the growth of CHO cells and a supplemental medium, wherein the combined basal medium and supplemental medium comprise an antioxidant, L-glutamine, iron, ethanolamine, and albumin; and insulin, wherein the insulin may be present in either the basal medium or the supplemental medium, or both. Optionally, the medium may contain EGF or IGF.

The invention relates to a serum-free adipose tissue-derived mesenchymal stem cell culture medium, which consists of a basic culture medium and added ingredients, wherein the basic culture medium is DMEM-LG, and the added ingredients and the content of each added ingredient are shown as follows: 5 to 20ng / mL of alkaline fiberblast growth factors, 5 to 20ng / mL of epidermal growth factors, 100U / mL of penicillin, 100 micrograms / mL of streptomycin, 50 to 200 micrograms / mL of heparin, 2 to 8mM of L-glutamine, 100 to 300 microM of 2-mercaptoethanol, 500 to 2000U / mL of leukaemia inhibitory factors and 0.5 to 2mM of sodium pyruvate. The serum-free adipose tissue-derived mesenchymal stem cell culture medium does not contain the serum, so that the inter-batch difference and the influence of the serum component on the cell culture can be avoided; the exogenous pollution of the serum and the toxicity of the serum on the cells can be avoided; and the ingredients are clear, so that the research of the psychological regulation mechanism of the cells can be facilitated.

The invention discloses an enrichment, domestication and screening method and application of an aerobic degradation microbial community of polycyclic aromatic hydrocarbon. An open glass bottle added with the polycyclic aromatic hydrocarbon and a basic medium is added with soil polluted by petroleum to enrich and domesticate aerobic degradation microbes of the polycyclic aromatic hydrocarbon; after30 days, a conical flask added with polycyclic aromatic hydrocarbon, a basic medium, trace liquid metal, vitamin c solution and an adsorbent is inoculated with the soil after the enrichment and domestication, and the soil is cultured in an oscillator at a temperature of between 25 and 30DEG C and a speed of 100r / min for 5 to 7 days, and then is subjected to circular transfer inoculation; and theaerobic degradation microbial community capable of highly efficiently removing the polycyclic aromatic hydrocarbon can be obtained after the circulation times is more than 8. The method can enrich, domesticate and screen the microbial community with good aerobic degradation performance on the polycyclic aromatic hydrocarbon, and aerobic degradation rates of the microbial community on naphthaline,phenanthrene, fluorine and pyrene are respectively 5.46mg.L<-1>.d<-1>, 0.48mg.L<-1>.d<-1>, 0.05mg.L<-1>.d<-1> and 0.06mg.L<-1>.d<-1>.