309 results about "HEPES" patented technology

Improved flow cytometer system particularly adapted to use for sex-selected sperm sorting include enhanced sheath fluid and other strategies which minimize stress on the sperm cells, including a 2.9 percent sodium citrate sheath solution for bovine species and a hepes bovine gamete media for equine species. Improved collection systems and techniques for the process are described so that commercial applications of sperms samples as well as the resulting animals may be achieved.

Improved insemination systems particularly adapted to use for sex-selected sperm sorting include systems which achieve superovulation and then multiple embryo production with sexed embryos. These systems combine with other techniques, including techniques for enhanced sheath fluid and other strategies which minimize stress on the sperm cells, and, potentially, a 2.9 percent sodium citrate sheath solution for bovine species and a hepes bovine gamete media for equine species. Improved collection systems and techniques for the process are described so that commercial application of sperms samples as well as the resulting animals may be achieved.

The invention discloses a fluorescence quantitative PCR detection kit of hepatitis B virus and an application thereof. The kit is composed of the following independent components: DNA extraction solution I, DNA extraction solution II, DNA extraction solution III, DNA extraction solution IV, positive control interior label, PCR reaction liquid, probe HBV-SP, enzyme mixed liquor containing heat resistant DNA polyase and uracil DNA glycosylase, quantitative hepatitis B virus reference material, hepatitis B virus positive control serum and hepatitis B virus negative control serum, wherein DNA extraction solution I contains 0.2-1.0% of lauryl sodium sulphate (mass/volume), 1.0-4.0% of Triton (volume/volume) and 0.2-1.0mol/L of guanidinium isothiocyanate; DNA extraction solution II contains 100-300mmol/L of 4-HEPES, 100-300mmol/L of sodium chloride with pH of 6.5+/-0.2 and 100-400 mu g/ml of magnetic beads; DNA extraction solution III contains 0.1-1.0% of Triton (volume/volume) and 100-300mmol/L of sodium chloride; DNA extraction solution IV contains mineral oil. The fluorescence quantitative PCR detection kit of hepatitis B virus of the invention can be used for detecting the HBV-DNA concentration in samples of serum, blood plasma or latex and the like.

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 phenanthrene and imidazole-coumarin double-fluorescent group ratio fluorescent molecular probe for iron ion detection and synthesis and use methods thereof, relates to fluorescent molecular probes and synthesis and application thereof and aims to solve the problem that an existing Fe<3+> fluorescent probe is prone to being interfered by pH, concentration and other metal ions. The fluorescent molecular probe is 4-methyl-7-hydroxide radical-8-[2-(1- phenyl group-1H-phenanthrene and [9, 10-d] imidazole-2-)benzene ammonia methylene]-2H-pyran-2-ketone. The phenanthrene and imidazole-coumarin double-fluorescent group ratio fluorescent molecular probe is formed by conducting condensation on 1-N-phenyl group-2-(2-aminophenyl)-1H-phenanthrene and [9, 10-d] imidazole and 4-methyl-7-hydroxide radical-8- formyl group coumarin, and the yield is 75-85%. The fluorescent molecular probe is dissolved in mixed liquid of N, N- dimethylformamide and an HEPES buffering solution, existence of iron ions is judged through the absorbance value or fluorescence intensity change before and after adding of test samples, and the fluorescent molecular probe can be used for detection of Fe<3+> pollution in water.

The invention discloses base fluid for diluting semens of equus animals and a preparation method and a use method thereof. The base fluid contains A fluid and skimmed milk. The A fluid contains glucose, raffinose, D-glucopyranose, trisodium citrate, ppotassium citrate and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES). The preparation method for the base fluid includes weighing various components of the A fluid into a sterile beaker, adding double-distilled water, dissolving, bringing to volume, filtering, split charging, sterilizing, and sealing to obtain the A fluid, and adding same amount of the skimmed milk into the A fluid to be evenly mixed. Users can also add penicillin and dihydrostreptomycin to the base fluid or add penicillin, dihydrostreptomycin, yolk and glycerol into the base fluid. The base fluid is suitable for semen dilution and normal temperature preservation, low temperature preservation and freezing preservation of the semens of the equus animals. The base fluid for diluting semens of equus animals and the preparation method and the use method of the base fluid have the advantages of being simple to prepare, convenient to use, good in preservation effect of the semens of the equus animals, multi-purpose, time-saving, labor-saving and applicable to mass production and application.

The invention discloses a specific culture medium for a lung tumor organ and a stentless 3D culturing method. The specific culture medium is prepared from the following components: FBS, double antibody, N-2, Noggin, B-27, EGF, FGF-10, Y-27632, A 83-01, SB202190, N-acetylcysteine, HEPES, Glutamax, IGF-1, hydrocortisone and Advanced DMEM/F12. The culturing method comprises the following steps: adding a tumor cell into a low serum culture medium, re-suspending the tumor cell, inoculating the tumor cell into a culture vessel, adding the specific culture medium into the culture vessel, changing thespecific culture medium once a day, and performing culturing until an organoid is formed. According to the culture medium and culturing method, a tumor organoid can quickly generate, can be stably cultured for a long time, is regular in spheroid form and has uniform and controllable size, and the heterogeneity of a tumor tissue of a patient can be well maintained in vitro.

Disclosed is a cornea middle term preserving fluid which comprises RPMI 1640 culture liquid, chondroitin sulfate, hyaluronic acid, HEPES cushioning liquid, dexamethasone, low-molecular-weight seaweed polysaccharides, Tobramycin or vancomycin, the preserving fluid has stabilized cell activity during storage period, and higher endothelial cell metabolizing function can be maintained than the US's Opthiol.

The invention relates to a tumor tissue tumor infiltrating lymphocyte (TIL) cell preparation method and a dedicated culture medium. The method comprises the following steps of tumor surrounding tissue obtaining, cell digestion, cell primary culture, cell subculture and cell collection, wherein a primary culture medium is based on a RPMI 1640 culture medium and prepared from the following concentration ingredients of 10% volume of human-derived serum, 20 to 45ng/ml of basic fibroblast growth factor (bFGF), 1 to 5mg/ml of riboflavin, 70 to 90ng/ml of cortisol, 10 to 25mg/ml of sodium dihydrogen phosphate monohydrate, 47 to 62ng/ml of recombinant human leukaemia inhibitory factor (LIF) and 500 to 800U/ml of IL-2; a subculture medium is based on the RPMI 1640 culture medium and prepared from the following concentration ingredients of 10% volume of the human-derived serum, 20 to 40mmol/L of HEPES, 1000 to 2000U/ml of the IL-2, 0.03 to 0.07mmol/L of beta-mercaptoethanol and 5 to 15ng/ml of sodium phosphate. According to the preparation method, an existing culture medium is improved, different culture mediums are utilized to culture the TIL cells in pertinence, TIL cell expansion capacity is improved, meanwhile a culture period is reduced, a culture complexity degree is reduced, a use amount of the IL-2 is reduced, and toxic reaction is reduced.

The invention discloses a fluorescein rhodamine B double-Schiff alkali compound ultraviolet molecular probe for Fe<3+> detection as well as synthesis and application thereof, and relates to a Fe<3+> probe as well as synthesis and application thereof. The technical problems that an apparatus required by an existing Fe<3+> fluorescent molecular probe is expensive and the ultraviolet probe is susceptible to interference are solved. A structural formula of the ultraviolet molecular probe disclosed by the invention is that the formula (1) is shown in the description. A preparation method comprises the following steps: carrying out condensation reaction between single-imino fluorescein aldehyde and rhodamine B hydrazine, and carrying out column chromatography separation on a mixed solvent of petroleum ether and ethyl acetate to obtain the ultraviolet molecular probe; dissolving the probe into a mixed solution of DMF (N,N-Dimethyl Formamide) and an HEPES (2-[4-(2-Hydroxyethyl)-1-Piperazinyl] Ethanesulfonic Acid) buffer solution; detecting ultraviolet spectral absorbance values before and after addition of to-be-detected samples, and judging that a sample solution contains Fe<3+> if the absorbance values of parts on which the wavelengths are 323nm and 358nm are increased. The method is simple and quick.

The invention relates to a reversible di-hydroxyl phenanthroimidazole Hg<2+> fluorescence probe and synthesis and use methods thereof, and relates to a fluorescence molecular probe and synthesis and application thereof. The reversible di-hydroxyl phenanthroimidazole Hg<2+> fluorescence probe provided by the invention has the advantages that the structure is novel; the technical problems of interference of the existing other metal irons on Hg<2+> recognition and the lack of reversible (off-on-off) fluorescence signal response detection are solved. The fluorescence molecular probe is formed by 1-N-1H-2-(2-hydroxyl-3-aminophenyl)-1H-phenanthro[9,10-d] imidazole and salicylaldehyde through condensation. The yield is 68 percent. When the fluorescence molecular probe is dissolved into a mixed solution of N,N-Dimethylformamide (DMF) and N-(2-Hydroxyethyl)piperazine-N'-2 ethyl sulfonic acid (HEPES) buffer solution, the fluorescence intensity is about 4 a.u.; after the Hg<2+> is added into a test sample, the fluorescence intensity is enhanced by 10 times; after the addition of S<2->, the fluorescence intensity recovers to the sample fluorescence value; the reversible di-hydroxyl phenanthroimidazole Hg<2+> fluorescence probe can be used for Hg<2+> and S<2-> pollution detection in water.

The invention discloses a coumarin schiff-base copper ion complex-based fluorescent probe for thiol as well as a preparation method and an application thereof. Coumarin hydrazide and 2,6-pyridinedicarboxaldehyde are used as raw materials for synthesis of a coumarin schiff-base derivative (a compound 1), the compound 1 and copper ions are coordinated in order to form the coumarin-copper ion complex with a schiff-base structure, and the compound 1-Cu<2+> is obtained. The probe ensures fluorescence quenching of the compound 1 due to influences of paramagnetism and chelation of Cu<2+>. When thiol-containing amino acids are added, fluorescence of the compound 1-Cu<2+> is enhanced. Other amino acids which do not contain thiols do not influence the fluorescence effects. UV-visible spectrum absorption and fluorescence spectrum show that the compound 1-Cu<2+> has good selectivity and sensitivity for thiol-containing amino acids in a CH3CN:HEPES(3:2,v/v) solution. The probe was successful to be applied to SiHa cells, and was used for carrying out cell imaging experiments of glutathione. The method is hopeful to be applied to detection of thiol compounds in organisms.

The invention discloses a serum-free culture medium suitable for immune cell large-scale culture, and belongs to the field of cell biology and medical immunology. The serum-free culture medium takes an RPMI1640 culture medium as a basis, additive components are further contained and include HEPES, phenol red sodium salt, L-glutamate and sodium bicarbonate. Insulin, transferrin, selenium, human albumin and interleukin-2 can be further added. Components contained in the culture medium are chemical defined components, no animal serum components are contained, and the serum-free culture medium is suitable for large-scale culture of immunity cells and is high in clinical safety and good in proliferation effect.

The invention puts forward a universal cancer organoid in-vitro culture medium, which consists of a DMEM/F-12K (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12K) basic culture medium and an adding factor, wherein the adding factor consists of HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), L-Glutamine, EGF (epidermal growth factor), Noggin, FGF (fibroblast growth factor)-10, A83-01 and Y27632. According to the culture medium of the embodiment of the invention, the success rate of organoid passage culture can be greatly improved, in addition, the universal culture of cancer samples, including gastric carcinoma, rectal carcinoma, lung carcinoma and the like, can be realized, culture cost is greatly lowered, the long-term culture of the organoid can be realized, and abiobank is established.

The invention relates to the fields of cytobiology and cell culture, and particularly relates to a breast cancer organoid culture kit. A method for realizing the kit comprises the following steps: (1)preparing, sub-packaging and using a breast cancer organoid culture medium A; (2) preparing, sub-packaging and using a breast cancer organoid culture medium B; and (3) preparing, sub-packaging and using a sample preserving fluid. The breast cancer organoid culture medium A is a DMEM/F12 culture medium containing 100 U/ml of penicillin, 0.1 mg/ml of streptomycin, 1% of HEPES and 1% of Gluta Max 100 X; the organoid culture medium B is a DMEM/F12 culture medium containing EGF, an FGF growth factor, an ALK inhibitor A83-01, Noggin, an HEPES buffer solution, Y27632 and one or more of a Wnt signalpath related secretory protein family R-Spondin 1-4; and the sample preserving fluid is a DMEM/F12 culture medium containing 100 U/ml of penicillin, 0.1 mg/ml of streptomycin and 1% of L-glutamine. The three components form the breast cancer organoid culture kit.

The invention provides a coating buffer for promoting cell adherence and a separation culture method of primary tumor cells. The coating buffer for promoting cell adherence comprises a type I collagen, a coating solution II and a coating solution III which are uniformly mixed, wherein the coating buffer II is a mixture containing 10*F12 medium, and the coating buffer III is a mixture composed of NaOH, NaHCO3 and HEPES. The scheme achieves separation culture of the primary tumor cells from smaller tissue samples.

The invention discloses a Chinese hamster ovary culture medium as well as a preparation method and application thereof. The culture medium comprises ascorbic acid, pyridoxine hydrochloride, vitamin B12, nicotinamide, riboflavin, thiamine chloride, choline chloride, folic acid, calcium pantothenate, sodium pyruvate, reduced glutathione, inositol, biotin, hypoxanthine, putrescine, lipoic acid, linoleic acid, thymine, glucose, HEPES (2-hydroxyethyl) buffer solution, dextran sulphate, Pluronic-F68, various amino acids and salts thereof and various inorganic salts. The culture medium has low cost, and the component does not contain animal origin ingredients, does not contain protein and has definite chemical ingredients. All ingredients are cooperated and blended, and the Chinese hamster ovary culture medium can satisfy the basic growth requirement of cells, and can effectively regulate, transfer and control the culture of cells so as to realize good high-density cell culture. Compared with the culture effect of the existing culture medium, the culture effect of culture medium disclosed in the invention is at least equivalent to even superior to the existing culture medium.

The invention discloses a preparation method of HEPES (hydroxyethylpiperazine ethane sulfonic acid) molecule guided porous zinc oxide microspheres. The preparation method comprises the specific experimental steps of taking HEPES molecules as a guide agent and organic zinc salt as a zinc source, reacting under a solvent heat condition at 150 DEG C, performing centrifugal washing to remove the residual solvent and HEPES modules, and drying the product to obtain porous zinc oxide microspheres with high specific areas. According to the preparation method disclosed by the invention, nontoxic HEPES molecules are used, the synthetic temperature is low, the operation is simple, and the repeatability is good. The prepared porous zinc oxide microspheres have the advantages of uniform size, multistage pore structure, large specific area and the like, and can be used as a photo-catalyst and a gas-sensitive element.

The invention discloses a preservation solution of a time-resolved fluorescent microsphere labeled myoglobin antibody. The preservation solution is prepared from the following components in percentage by mass: 0.1 percent to 1.5 percent of 4-(2-hydroxyethyl)piperazine-1-erhanesulfonic acid, 1.0 percent to 3.0 percent of sodium chloride, 0.3 percent to 0.8 percent of potassium chloride, 0.08 percent to 0.9 percent of disodium ethylene diamine tetraacetate, 0.01 percent to 0.06 percent of sodium dodecyl sulfonate, 0.1 percent to 3.0 percent of bovine serum albumin, 0.05 percent to 0.5 percent of glucan, 0.02 percent to 0.2 percent of triton, 0.01 percent to 0.05 percent of Tween-20, 0.02 percent to 0.1 percent of a preservative and the balance of water; the pH (Potential of Hydrogen) is adjusted to be 8.0 to 9.0. The preservation solution has the advantages that the raw materials are low in cost and easy to obtain, and a relatively stable microenvironment is provided for preservation of fluorescent microspheres.

The invention discloses a combined medium for preparing insulin-like secreting cells, which consists of a medium I, a medium II and a medium III which are independent respectively, wherein, the medium I is a DMEM (Dulbecco's modified Eagle's medium) high-glucose medium prepared from fetal calf sera, basic fibroblast growth factors (bFGF), dimethyl sulfoxide (DMSO) and glucose; the medium II is a serum-free DMEM/F12 medium prepared from glucose, niacinamide, epidermal growth factors (EGF), bFGF, exendin-4, a B27 additive and an N2 additive; and the medium III is an RPMI (Roswell Park Memorial Institute) 1640 medium prepared from glucose, niacinamide, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, hepatocyte growth factors (HGF), activin A and exendin-4. The invention further discloses a method using the combined medium for preparing insulin-like secreting cells at the same time. The invention can facilitate the differentiation of hBM-MSC (human bone marrow mesenchymal stem cell) into insulin-like secreting cells having the effect of correcting diabetes hyperglycemia.