18 results about "Alanyl glutamine" patented technology

The invention provides an induced pluripotent stem cell culture medium and a culture method of induced pluripotent stem cells. The induced pluripotent stem cell culture medium comprises a basic culture medium, a composite additive A and a composite additive B, the composite additive A comprises 5-30 [mu] g / mL of recombinant human insulin growth factors, 1-5 ng / mL of recombinant human transforming growth factors, 50-200 ng / mL of recombinant human basic fibroblast growth factors, 7-20 [mu] g / mL of sodium selenite, 0.5-2 mg / mL of albumin, 50-70 [mu] g / mL of ascorbic acid and 5-20 [mu] g / mL of transferrin; the composite additive B is prepared from 50 to 300mg / mL of a Stock B solution, 0.5 to 5 percent (v / v) of a non-essential amino acid solution, 50 to 200ng / mL of DL-piperidinecarboxylic acid, 50 to 200mu g / mL of gamma-aminobutyric acid, 0.1 to 1 percent (v / v) of a lipid concentrated solution, 0.5 to 5 percent (v / v) of an L-alanyl-L-glutamine solution and 0.1 to 0.1 percent of mercaptoethanol. The culture medium for the induced pluripotent stem cells provided by the invention can ensure the self-renewal capability of the iPSCs in a long-term passage process; and the genetic stability of the iPSCs cells is optimized according to the genome integrity and pluripotent state.

The invention discloses a method for synchronously detecting L-asparagine monohydrate and L-alanyl-L-glutamine in an assisted reproduction culture solution. The L-asparagine monohydrate and the L-alanyl-L-glutamine in the assisted reproduction culture solution are used as a detection reagent. According to the method, a pretreatment mode of combining protein precipitation with diluted hydrochloric acid extraction is adopted, a target object is separated through ion exchange chromatography, and a ninhydrin post-column derivatization technology is adopted to establish an external standard method quantitative analysis system. The method has the following advantages: (1) synchronous and efficient separation and accurate quantification of two amino acid substances are realized; (2) methodology verification shows that good repeatability (RSD (Relative Standard Deviation) is 1.05) and high sensitivity (the detection results of L-asparagine monohydrate and L-alanyl-L-glutamine are 0.00038 mg / mL and 0.00036 mg / mL respectively); (3) the operation is simple and convenient, and the single analysis time is about 60 minutes; and (4) the recovery rate is stabilized in a range of 93%-105%. The method provides a reliable analysis means for quality control of the assisted reproduction culture solution, and is particularly suitable for quantitative determination of key amino acid components in the assisted reproduction culture solution.

The present application belongs to the technical field of feed additive, and discloses a kind of feed additive for improving the quality of Nibea albipunctata swim bladder, which is composed of the following components by mass fraction: 40-55wt% cottonseed protein, 25-38wt% soybean saponin, 10-16wt% capric acid monoglyceride, 8-10wt% alanyl glutamine. In the above-mentioned feed additive, alanyl glutamine and capric acid glyceride play an important role in the process of synthesizing swim bladder collagen protein and accumulating essential amino acids in collagen protein. The mutual cooperation between the components in the feed additive can promote the deposition of swim bladder collagen protein and improve the accumulation of essential amino acids in collagen protein at the same time.

The invention relates to methods for extracting, purifying and concentrating mesenchymal stromal cell exosomes enriched with micro-RNA, proteins and lipids, and can be used in cosmetology and pharmacology for rehabilitation and regeneration. The technical result of the invention is a more efficient process for producing exosomes. This technical result is achieved in that the present method for extracting, purifying and concentrating exosomes from mesenchymal stromal cells includes cultivating cells, collecting the conditioned culture medium, depleting high-molecular-weight proteins, extracting, purifying and concentrating exosomes, and freezing and lyophilizing the exosome concentrate; in order to enrich the culture medium with exosomes, multipotent mesenchymal stem cells (MSCs) are grown in a medium having a lowered glucose concentration and containing platelet lysate and 5 mM of L-alanyl-L-glutamine, wherein, in the case of static cell cultivation, the growth medium is placed in a culture vessel and once a level of not less than 80% monolayer confluence is reached, the culture medium is harvested and replaced once every 24 hours, and in the case of flow-through cell cultivation, cells are placed on cell carriers in a bioreactor and 72-96 hours after the start of cultivation, the culture medium is harvested and replaced once every 24 hours, the exosome-enriched culture medium is subjected to concentration, which includes successive stages of centrifugation, tangential flow filtration and isopycnic centrifugation: in the first stage, the culture medium is centrifuged for 10 mins at 2000 g to remove large cell particles and other inclusions; in the tangential flow filtration stage, the supernatant is concentrated using hollow fibre filters having a molecular weight cutoff of 500 kDa, after which the concentrate is subjected to fivefold diafiltration with an equal volume of DPBS phosphate buffer solution using the same hollow fibre cartridge; and in the isopycnic centrifugation stage, the concentrated supernatant is mixed with a saccharose solution and deuterium oxide and subjected to isopycnic centrifugation at 100000 g and 4˚С for 75 mins, after which the concentrate is subjected to further fivefold diafiltration with a buffer solution, then the concentrate is filtered through filters with a pore diameter of 0.22 µm and 0.1 µm, the exosome concentrate is subjected to initial freezing to -75˚C in a freezing chamber, and drying is carried out in a lyophilization chamber, wherein in a first drying stage, the concentrate is cooled to -40˚С and subsequently held for 100 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -30˚С and subsequently held for 610 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -10˚С and subsequently held for 315 mins at an absolute pressure in the chamber of not more than 10 Pa, after which it is cooled to 0˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa, then heated to +10˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa; in a second drying stage, drying is carried out at +22˚С for not less than 130 minutes at an absolute pressure in the chamber of not more than 10 Pa.

The invention belongs to the technical field of biological medicines, and discloses an endometrial organ and a construction method thereof. The invention provides a construction method of endometrial organs. An IM differential culture medium 1 containing L-alanyl-L-glutamine with specific concentration and CHIR 99021, an IM differential culture medium 2 containing L-alanyl-L-glutamine with specific concentration, bFGF and RA and an MD differential culture medium containing CHIR 99021 with specific concentration are adopted as a Miller tube cell induced differentiation culture system; the method can efficiently guide the differentiation of the human pluripotent stem cells to the Mullerian canal cells while maintaining the cell proliferation ability, the obtained Mullerian canal cells have excellent proliferation and differentiation ability, and an organoid with natural endometrial characteristics can be formed in an endometrial organoid induced differentiation culture system; the material can be used as an experimental material in research on development mechanism of endometrium, disease model establishment, drug screening and the like, and has a wide application prospect.

The invention discloses an embryo culture solution as well as a preparation method and application thereof, and belongs to the technical field of assisted reproduction. The technical problem to be solved is to reduce the ammonium ion concentration in the use process of the cell culture fluid. According to the technical scheme, the embryo culture solution is characterized by comprising sodium chloride, potassium chloride, monopotassium phosphate, calcium chloride, magnesium sulfate, sodium bicarbonate, a sodium lactate solution, sodium pyruvate, glucose, edetate disodium, gentamicin sulfate, human serum albumin, taurine, alanyl glutamine, non-essential amino acid and essential amino acid. The total concentration of the alanyl glutamine, the non-essential amino acid and the essential amino acid is 0.3-0.7 g / L; the mass ratio of the alanyl glutamine to the non-essential amino acid to the essential amino acid is (0.05 to 0.2) to (0.03 to 0.05) to (0.2 to 0.45); the mass ratio of the human serum albumin to the alanyl glutamine is (3-5): (0.05-0.2).

A complete culture medium for efficient culture and amplification of human umbilical cord mesenchymal stem cells is characterized by comprising the following components: a basic culture medium accounting for 85%-93% of the complete culture medium; the addition proportion of the UltraGRO accounts for 3%-6% of that of the complete culture medium; the addition proportion of the KnockoutTM SR accounts for 3%-6% of that of the complete culture medium; the addition proportion of the L-alanyl-L-glutamine accounts for 1%-3% of that of the complete culture medium. According to the efficient culture and amplification method for the human umbilical cord mesenchymal stem cells, the complete culture medium for efficient culture and amplification of the human umbilical cord mesenchymal stem cells is used. When the complete culture medium with the components and the proportion is used for efficiently culturing and amplifying the human umbilical cord mesenchymal stem cells, the cell morphology of the human umbilical cord mesenchymal stem cells is still not obviously changed until the cell morphology of a P10 generation, the amplification multiple is not obviously changed after being stably passed to the P10 generation, the human umbilical cord mesenchymal stem cells are stably amplified, and the HGF secretion amount is also obviously increased; obviously, the culture amplification effect is obviously improved.

This document relates to compositions, methods, and formulations for evaluating and / or treating subjects (e.g., humans) having ophthalmic diseases, disorders, and pathological conditions. For example, the methods and compositions disclosed herein include ophthalmic pharmaceutical compositions comprising L-alanyl-L-glutamine suitable for the eye.

The present application belongs to the field of biotechnology, and relates to a method for producing coxsackie virus by using HeLa cells. Specifically, the present application relates to a method for culturing HeLa cells in vitro to produce coxsackie virus, which comprises: sequentially culturing HeLa cells in culture medium III and culture medium I, inoculating virus and culturing to obtain a cell culture containing coxsackie virus; wherein the culture medium III is a mixture of culture medium II and VirusPro culture medium at a volume ratio of 3:1-1:1, the culture medium I is a mixture of culture medium II and VirusPro culture medium at a volume ratio of 3:1-1:3, and the culture medium III and the culture medium I contain L-alanyl-L-glutamine.

The invention relates to the technical field of bioengineering, in particular to a culture medium and a method for constructing auditory neuroma organs. According to the culture medium, Neurobase Medium and DMEM (Dulbecco Modified Eagle Medium) / F12 are used as a basic culture medium; the feed additive comprises a N2 additive, a B27 additive, L-alanyl-L-glutamine, an antibiotic, HEPES, glucose, beta-mercaptoethanol, a cell growth factor, MEM NEAA and a TGF-beta inhibitor. The cell growth factor is EGF (Epidermal Growth Factor) or FGF-2. The culture medium provided by the invention can effectively improve the construction efficiency of the auditory neuroma organoid, and the constructed auditory neuroma organoid can be used as a reliable in-vitro substitution model, is used for marker detection, drug screening and individualized medical exploration in basic research and clinical transformation research, and has important application value.

This invention discloses an amino acid assay kit based on the o-phthalaldehyde (OPA) method, its preparation method, and its application. Belonging to the field of amino acid detection and analysis technology, it is used for rapid and accurate quantitative detection of amino acids in samples. The amino acid assay kit can not only measure the total amino acid content but also analyze specific amino acids and dipeptides such as alanine glutamine, meeting diverse analytical needs. This amino acid assay kit is suitable for batch testing and supports high-throughput laboratory applications. By optimizing the reagent formulation and operating procedures, this invention effectively simplifies the detection steps, improves detection sensitivity and specificity, and fills the gap in the market for OPA-based assay kits. This invention can be widely applied in nutrition, medicine, clinical diagnostics, and drug development, providing a convenient and efficient amino acid analysis tool for life science and medical research.

The application provides a high performance liquid chromatography method for determining related substances in alanyl glutamine bulk drug, adopts a chromatographic column with octadecylsilane bonded silica gel as a filler, can effectively separate alanyl glutamine related substances, including degradation impurities and process impurities, and the method has high precision, long service life of the column, and can better control the product quality of the alanyl glutamine bulk drug.

The application relates to the field of biological medicine, and particularly relates to a cold storage solution and application of a combined traditional Chinese medicine compound biological stent drug delivery system in eardrum repair. The cold storage solution comprises a basic solution and L-alanyl-L-glutamine; the basic solution comprises a STEEN solution. The cold storage solution can promote metabolism and synthesis of glutathione of fat grafts, regulate ROS mitochondrial oxidative stress, reduce cold storage damage of the fat grafts, and improve the cold storage quality of the fat grafts. Meanwhile, the application uses Shengji Yuhong collagen sponge which contains an important classical prescription for promoting repair, forms a functional biological material with supporting and pro-angiogenic effects, forms a linkage mode of "double-stage regulation before and after transplantation", closely combines the actual difficulties of cell damage of fat groups and reduction of repair success rate caused by long eardrum repair time in the clinic, explores a brand-new research mode for improving the eardrum repair scheme, and has far-reaching clinical significance.

The invention belongs to the technical field of biology, and relates to a method for producing CVB1 virus by culturing HEK293 cells in vitro, which comprises the following steps: (1) culturing host cells in a culture medium I which comprises a basic culture medium of L-alanyl-L-glutamine; (2) adding a culture medium II into the cell culture in the step (1), and continuously culturing the host cells; (3) adding a culture medium III into the cell culture in the step (2), inoculating viruses, and continuously culturing the host cells inoculated with the viruses; and (4) obtaining a cell culture containing the target virus. The invention also relates to culture media for the production of CVB1 viruses, and their use for the production of CVB1 viruses.

The utility model relates to a rotary evaporator for alanyl glutamine production, which comprises a base, the top of the base is fixedly provided with a water bath kettle, the right side of the water bath kettle is hinged with a protective cover, the top of the base is fixedly provided with a support seat positioned on the left side of the water bath kettle, the inside of the support seat is fixedly provided with a lifting component, and the lifting component is fixedly provided with a lifting handle. A rotating device is fixedly mounted on the front side of the lifting assembly, a conveying pipe is fixedly mounted in the rotating device, and under the action of a first damping spring telescopic rod, a second damping spring telescopic rod and a buffer plate, the influence of vibration generated in the working process of a motor at the rotating device on the part, connected with a condenser and an evaporation bottle, of the conveying pipe is reduced; and when the induction block reaches the position of the induction sensor, the lifting assembly stops working, collision is avoided, the periphery of the water bath kettle can be protected through the protective cover during working, and the influence of air convection on the evaporation efficiency is reduced.

The invention relates to a cell freezing medium as well as a preparation method and application thereof. The cell freezing medium contains polyvinyl alcohol, taurine, serine, L-alanyl-L-glutamine, L-carnitine, dextran, trehalose, poloxamer 407, vitamin C magnesium phosphate, DMSO (dimethylsulfoxide) and a cell basal culture medium. The invention provides the serum-free and protein-free cell cryopreservation liquid limited by chemical components, the components are clear and stable, the problem of unstable cryopreservation effect caused by complex components and inter-batch difference is avoided, the pollution risk and immunogenicity are reduced, and the serum-free and protein-free cell cryopreservation liquid is particularly suitable for preserving target cells for clinical treatment.

The application discloses a kind of cell spheroid serum-free cryopreservation solution and its preparation method and application, steps are mixed to complete mixing with a certain mass of DMSO, ethylene glycol and glycerol, as composite osmotic protection phase;A certain mass of methyl cellulose, trehalose and hydroxyethyl starch are added in a certain volume of ultrapure water and stirred to dissolve, then a certain mass of cholesterol, Y27632 and glutathione are stirred to completely dissolve, as non-osmotic protection phase;A certain mass of composite osmotic protection phase and non-osmotic protection phase are mixed, and a certain mass of KOSR serum substitute, L-alanyl-L-glutamine and insulin is added, slowly stirred and filtered to remove bacteria, to obtain cell spheroid serum-free cryopreservation solution.The cell spheroid cryopreservation solution prepared by the application combines low-toxicity composite osmotic protection and multiple non-osmotic protection, realizes high recovery survival rate and stable structure maintenance, and the process is simple, which is very suitable for the cryopreservation of multiple types of cell spheroids.

The invention provides an optimized umbilical cord mesenchymal stem cell culture solution and a preparation method thereof, and relates to the technical field of cell culture, the optimized umbilical cord mesenchymal stem cell culture solution comprises a basic culture medium, a platelet lysate, a cell generation factor, L-alanyl-L-glutamine and a mixed solution; the mixed solution consists of an antioxidant solution, an adherent proliferation solution and a metabolic regulation solution. The selected basic culture medium can provide an adaptive basic nutrition environment for cells, the blood platelet lysate is matched to reduce exogenous pollution, and meanwhile, the blood platelet lysate and the cell generating factors have a synergistic effect, so that efficient proliferation of the cells is assisted, the dryness of the cells can be stably maintained, and the loss of a dryness marker in the passage process is avoided; by adding the L-alanyl-L-glutamine, the problem of ammonia toxicity of conventional glutamine can be avoided, stable cell metabolism is guaranteed, and the mixed solution can optimize a cell growth microenvironment from multiple aspects, assist cells to keep a good growth state, ensure high activity of the cells and reduce the risk of cell apoptosis.