66 results about "Cell metabolite" patented technology

Embodiments of the present invention relate to analyzing components of a cell. In some embodiments, the present invention relate to analyzing components of a single cell. In some embodiments, the methods and compositions relate to sequencing nucleic acids. In some embodiments, the methods and compositions relate to identifying and / or quantitating nucleic acid, proteins, organelles, and / or cellular metabolites.

A three-dimensional (3-D) culture “Petri-dish” for research in regenerative medicine, biotechnology and clinical translation is described. This 3-D perfusion culture dish is to advance in vitro culture tools from static 2-D to dynamic 3-D perfusion culture. Interwoven hollow fiber capillary membranes divide the “Petri-dish” culture space into a controllable 3-D pattern of different compartments, serving the functions of the organ's larger vasculature. These physically active scaffolds, which can be suitable for cell adhesion or cell aggregate immobilization, offer a supply of cells with high-performance mass exchange including gas supply and under perfusion conditions. In contrast to static and discontinuous medium supply, a dynamic culture can be achieved with continuous or alternating medium supply and integral oxygenation. They provide a more physiologic supply in the cell macro environment, including homeostasis of oxygen, pH, nutrition, soluble factors, and gradients of metabolites for the cells. Also, medium perfusion can be achieved. Consequently the invention was made for cultures at tissue density, especially stem cells and support cells, which strive to create their own stem cell niche.

The invention provides biomarker profiles of cellular metabolites and methods for screening chemical compounds including pharmaceutical agents, lead and candidate drug compounds and other chemicals using human embryonic stem cells (hESC) or lineage-specific cells produced therefrom. The inventive methods are useful for testing toxicity, particularly developmental toxicity and detecting teratogenic effects of such chemical compounds.

The invention relates to an isolated organ preserving device and a preserving method thereof, and belongs to the technical field of medical equipment. The isolated organ preserving device is structurally characterized in that continuous mechanical irrigation and membrane oxygenation are adopted, an isolated organ continuously obtains nutrients and the oxygen supply, a cell metabolite is removed in time, and a cell microenvironment is better maintained, so that the function normality is ensured. The isolated organ preserving device disclosed by the invention can be used for directly contacting the organ perfusate and the organ perfusion instrument by adopting a disposable consumable material sterilized through epoxyethane, thereby meeting the requirement for sterility of a system. Besides, the isolated organ preserving device disclosed by the invention can be used for judging the consumption of the nutrients contained in the organ perfusate and the gathering degree of metabolic wastes by monitoring the pH value of the organ perfusate in real time and can be used for abandoning the organ perfusate after a certain standard is achieved and replacing the fresh organ perfusate, thereby further ensuring the possibility of long-time preservation; the isolated organ preserving device can be used for evaluating the damage condition of the isolated organ by monitoring the front and back perfusion pressure of the isolated organ in real time and converting the perfusion resistance by combining a flow number.

The invention discloses a multichannel microfluidic-solid phase extraction-mass spectrum combined device and a preparation method. The combined device comprises a microfluidic chip and a mass spectrometer, wherein the microfluidic chip is provided with upper and lower layers of channels; the upper layer of channel is a cell culture channel; the two ends of the upper layer of channel are provided with a vertical hole and a horizontal hole respectively; the lower layer of channel is a chemical oxygen uptake channel; the two ends of the lower layer of channel are provided with a plurality of groups of vertical holes respectively, and the upper and lower layers of channels are connected through a layer of PDMS (Polydimethylsiloxane) membrane; C18 spherical particles are filled on one side of the upper layer of channel to form a filling column; the horizontal hole of the upper layer of channel is connected with the mass spectrometer. The physiological behaviors of cells under different oxygen environments can be simultaneously observed, and control experiment, multi-concentration gradient parallel stimulation and multi-region detection can be realized at one time. Through separation and enrichment of the solid-phase extraction chip, mass spectrum real-time detection is realized, cell metabolites can be more intuitively and more accurately detected, and a theoretical basis is provided for research of tumor cell hypoxia.

Methods for treating diseases in humans and vertebrate animals are provided using competitive antagonists of cellular metabolites combined with a protective agent for protecting host cells from toxic effects of the drugs. Also provided are kits comprising competitive antagonists and suitable protective agents. In addition, screening methods for identifying competitive antagonists, protective agents and potentiating agents, for use according to the methods of the invention, are provided.

The invention discloses a new method for performing data correction by using cell metabolite relative content as cell number index. The new method is characterized by comprising the steps of: realizing the fast quenching of biochemical metabolism in cells by using a simple sample pre-processing method; efficiently extracting small molecule metabolites by an optimized extraction solvent; and roundly and half quantitatively measuring the small molecule compounds in a biological sample by a metabonomics method based on measuring technology like mass spectrum, nuclear magnetic resonance and the like so as to identify a quantitative index of symbolic metabolites in cells to be as the cell number, and regarding the quantitative index as a weight factor to perform data correction. Compared with the quantitative index of the prior cell number, the method is simple, fast and with good repeatability; additional experimental working and the error caused by the additional working are reduced; the accuracy and reliability of experiment result are improved; the difficulty for relatively quantifying the number of in vitro cultured cells is solved; a new displacing method is supplied for cell metabonomics and relevant researches; and the method can be widely used in the relevant researches in the field of cell molecular biology.

The invention provides a proliferation culture medium for 3D simulation culture of stem cells, the proliferation culture medium is free of serum addition, definite in composition and suitable for 3D simulation perfusion culture of stem cells, and comprises inorganic salt, amino acid, saccharides, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors and water, the osmotic pressure of a culture solution is 270-330mOsmol / kg.H2O, and the pH value of the culture solution is 7.2-7.4.

The invention relates to a proliferation culture medium for 3D simulation culture of stem cells, the proliferation culture medium is free of serum addition, clear in composition and suitable for 3D simulation perfusion culture of stem cells, and comprises inorganic salt, amino acid, saccharides, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors and water, the osmotic pressure of a culture solution is 270-330mOsmol / kg.H2O, and the pH of the culture solution is 7.2-7.4.

The invention discloses a bone repair material with a multi-dimensional channel structure. The bone repair material has a layered matrix structure; a plurality of bosses I are formed in one surface of the matrix structure, and a plurality of bosses II are formed in another surface of the matrix structure; the bosses I and the bosses II are arrayed alternatively; grooves are formed between adjacent bosses I and adjacent bosses II; and a plurality of through holes are formed in a way of being perpendicular to the surfaces of the bosses I. The bone repair material provided by the invention adopts a three-dimensional network channel formed through the through holes in the matrix structure and the grooves, and the three-dimensional network channel is filled with collagenous fiber, which is favorable for cell attachment and growth as well as nerve ingrowth, and also is convenient for nutrient substance transfer and cell metabolite discharge, and also can enable cells to be evenly distributed in the interior of the material, and is favorable for even and uniform organizational formation in the interior of a support; the structural material can form firm biological gomphosis with regenerated new bone, and good structure stability can be maintained during a degradation process; the mechanical strength is high, and mechanical transmission between a bone defect part and normal bone tissue can be well achieved.

The invention discloses a method of screening a marker of acute renal toxicity caused by aristolochic acid by utilizing cell metabolic profiling in vitro. The effective dose causing renal cell damage is determined by adopting a 3-(4,5-dimethylthiahiazo-2)-2,5-diphenytetrazoliumromide (MTT) staining manner. Renal cell metabolites are analyzed by adopting an ultra-high performance liquid chromatography-mass chromatography technique to obtain metabolic profiles. Metabolic profile data of normal groups and medicine damage groups are analyzed by adopting a multivariate statistic method. A potential renal toxicity marker is screened by combining a change rule of metabolites along with medicine acting time. The screened marker is good in predicting performance. The average accuracy of outer verification is 90% or above. The method comprehensively represents variation situations of metabolites of normal renal cells under medicine damages, and can provide technique supports for early diagnosis of renal toxicity caused by medicines or medicine safety evaluation.

The invention provides a real-time cell metabolite detection device based on a micro-fluidic chip. The device comprises the micro-fluidic chip, an automatic sampling assembly, a pretreatment and enrichment assembly and a detector. Channels provided with cell culture rooms are arranged in the micro-fluidic chip, the automatic sampling assembly capable of conducting sampling at outlets of the channels is arranged around the micro-fluidic chip, and the pretreatment and enrichment assembly is connected to the down stream of the automatic sampling assembly and can conduct desalting and target product enriching on a cell metabolite obtained through sampling. The detector is arranged at the portion corresponding to an outlet of the pretreatment and enrichment assembly to conduct online detection on the target product. According to the detection device, the micro-fluidic chip, automatic sampling, pretreatment and enrichment and detection are integrated into a whole, and real-time online detection and analysis can be conducted on a biochemical reaction on the micro-fluidic chip and the drug metabolism process.

The invention relates to a method for analyzing influence of a multi-dimensional nano material on cell metabonomics. The analysis method for the influence of the multi-dimensional nano material on thecell metabonomics based on the liquid chromatography-mass spectrometry technology comprises the following steps: (1) co-culturing the multi-dimensional nano material and cells; (2) performing metabolite extraction; (3) carrying out liquid chromatography-mass spectrometry combined detection; and (4) performing bioinformatics statistical analysis. According to the metabonomics analysis method, theinteraction between the multi-dimensional nano material and cells is investigated, the cell metabolite is detected and analyzed, and the cell metabolite variety contents and the metabolic pathways under the effects of different multi-dimensional nanometer materials are compared so that the influence principle of the nanometer materials with different dimensions on the cell metabonomics is revealed, the basis is provided for the evaluation of the biological characteristics of the nanometer materials, and the application prospect is broad.

The invention discloses an in-vitro cell contact type co-culture device and a culture operation method thereof. According to the device, a box bottom is arranged at the bottom end of a culture box body; a box cover is arranged at the upper end of the box body; a cell culture tank is arranged in the culture box body; the culture box body is in the shape of a conical gyro, and the diameter at the upper end of the culture box body is larger than the diameter of the bottom of the culture box body; the cell culture tank is barrel-shaped; the upper and lower diameters of the barrel are the same as the diameter of the box bottom of the culture box body; an isolating membrane is fixedly arranged in the middle of the culture tank, and divides the culture tank into an upper culture tank and a lower culture tank. The culture tank is ingeniously divided into the upper culture tank and the lower culture tank by using the polycarbonate membrane and is respectively inoculated on the upper and lower sides, and the culture tank is arranged in the cell culture box after being overturned, so that substantive cell contact type co-culture is realized, and influence of metabolites of one kinds of cells on another kinds of cells and an intercellular effect caused by direct contact are conveniently researched.

Volatile organic compounds emitted from gastric cancer cell metabolite is separated and detected using HS-SPME / GC-MS. A volatile compounds fingerprint atlas-spectrum model are disclosed for early gastric cancer diagnosis / warning Volatiles 3-octanone and 2-butanone as indicators of gastric cancer cells which are not contained in the headspace of gastric mucosal cells GES-1. Meanwhile, the ratio of mass to volume to concentration of 4-isopropoxy alcohol, nonanoic acid, and 4-butoxy n-butanol is as follows: 4-isopropoxy alcohol [gastric cancer cells] / [normal gastric mucosa cells]≦0.31; nonanoic acid [gastric cancer cells] / [normal gastric mucosa cells]≦0.36; and 4-butoxy n-butanol [gastric cancer cells] / [normal gastric mucosa cells]≦0.40. The volatile organic compound in cell metabolite to be detected is compared with the fingerprint atlas-spectrum model, so as to implement screening and warning of the early gastric cancer.

The invention discloses use of bacillus coagulans JA845 in preparing an enzyme activity inhibitor with a hypoglycemic function and relates to the field of hypoglycemic drugs. The bacillus coagulans JA845 is prepared into a fermentation supernatant, a cell-free extract and a bacterial cell metabolite, activity inhibition rates of the bacillus coagulans JA845 to alpha-glucosidase and DPP-4 are measured. Results show that a to-be-detected sample of the bacillus coagulans JA845 has an activity inhibition effect on the alpha-glucosidase and the DPP-4, the cell-free extract of the bacillus coagulans JA845 had the highest inhibition activity, such that the bacillus coagulans JA845 and / or derivatives thereof can be used for preparing the activity inhibitor of the alpha-glucosidase and the DPP-4 and a new thought is provided for preparing hypoglycemic drugs.

The invention discloses an isolated culture method for tea leaf suspension single cells. The method comprises the following steps: selecting tea leaves; disinfecting aseptic leaves; inducing healing tissues; culturing liquid suspension cells; performing enlarged culture on the liquid suspension cells; separating the single cells and the like. The tea leaf suspension cells prepared by the method have high growing speed and higher metabolism activity. A rapid-growing suspension cell culture system and a single cell separating method are provided, so that a basis is laid for tea leaf cytological studies and researches of secondary metabolites. A good technical platform is provided for subsequent production of tea tree secondary metabolites, metabolite cellular localization, metabolic mechanism study as well as tea tree cell metabolite change and mechanism study under the stress condition of exogenous compounds.

The invention provides a method for culturing a suspended cell virus capable of maintaining the production stability of virus antigens and improving the effective content of viruses. The method uses aserum-free culture method in a cell culture stage, and the first-time medium change reduces the influence of cell metabolites on cell growth and reduces the interference on late-stage virus production. After the medium change, the added dextran sulfate effectively plays the effects of improving virus infection sensitivity and cell dispersion. PEG 6000 and a high-calcium solution are added to a later-phase virus inoculation and medium change system so as to improve the virus inoculation efficiency and to greatly increase the stability of foot-and-mouth disease virus. The titer of the preparedvirus solution can reach 107.0-109.01 ogTCID50 / mL, the content of complete antigen 146S can reach 6 [mu]g / ml or above, and the cultured foot-and-mouth disease virus is more stable.

The invention discloses a method for evaluating toxicity effects of low-dose combined exposure of organic pollutants based on a metabonomics technology. Firstly, single exposure and combined exposureare performed to HepG2 cells by two organic pollutants, unbiased and accurate metabolite information can be obtained based on quasi-targeted metabolomics analysis, and multiple comparative analysis and enrichment analysis of metabolic pathways can directly reveal the impact of the organic pollutants on cellular metabolites and the metabolic pathways. By calculation of the metabolic impact level index, the combined exposure effect type of the combined exposure on overall metabolism and specific metabolic pathways can be quantitatively described. A biological detection method can further determine the activity of metabolic pathway rate-limiting enzymes, and confirm the change direction of the metabolic pathways and the main risk-driven compounds in a mixture. The method has the advantages ofeasy operation, fast detection, high sensitivity, strong selectivity, good repeatability and obtaining of systematic and comprehensive information.

The invention provides a method for improving heterotrophic microbial fermentation via multiple chemical promoters to produce docosahexenoic acid (DHA). The method comprises the following steps: with a heterotrophic culture medium of crypthecodinium cohnii as a foundation, adding any chemical promoter into the heterotrophic culture medium; the heterotrophic culture medium is composed of 9g / L of glucose, 2g / L of yeast extract powder and 25g / L of sea salt; the chemical promoter can also be called a plant growth regulator, including indoleacetic acid (IAA), naphthoxyessigsaeure (BNOA), chlorobenzoic acid, salicylic acid (SA), abscisic acid (ABA) and cholamine, and the concentrations of these chemical promoters in the heterotrophic culture medium are as follows: 2.5mg / L of indoleacetic acid (IAA), 4mg / L of naphthoxyessigsaeure (BNOA), 4mg / L of chlorobenzoic acid, 1mg / L of salicylic acid (SA), 20mg / L of abscisic acid (ABA) and 2.5mM cholamine; culturing at 25 DEG C for 72 hours, extracting grease, and measuring the DHA content of the culture by using a gas chromatograph-mass spectrometer, meanwhile, extracting cellular metabolite from a parallel culture, and detecting and analyzing the content change condition of the cellular metabolite by using a metabonomics method.

Container equipment which can integrate an animal cell culture reactor and a separator is applied in experimental and production processes of animal cell culture and animal cell's metabolite extraction in the field of bio-fermentation engineering. The equipment is composed of an external tank with an elevating wall for adjusting volume and an internal tank with a fixed volume, and two work systems, namely an agitatory animal cell culture reactor and a separator, are configured. When the external tank and the internal tank are combined, the formed inner space can be used as a reactor to support the cell culture fermentation process under the control of an automation procedure. When the elevating wall is lifted by a hydraulic device, a separate disengagement chamber formed at the bottom of the tank is used to support the separation and extraction process under the control of the automation procedure. By the adoption of the equipment, cell metabolite can be extracted timely, continuously and conveniently, hygienic requirements can be guaranteed, and production efficiency can be raised.

The invention provides application of Cordyceps fungal cell metabolites in the preparation of skin-care products and a skin-care product; Cordyceps fungi herein at least include any one of Cordyceps sinensis, Hirsutella sinensis, Paecilomyces hepiali, Hirsutella hepiali, Cordyceps militaris, Cordyceps algae, Cordyceps ophioglossoides, Cordyceps hawkesii, Cordyceps martialis, Ophiocordyceps sobolifera, and Cordyceps fruit flies. The Cordyceps fungal cell metabolites herein are not typical antibiotics, but provide germ-killing action; the Cordyceps fungal cell metabolites are not typical anti-inflammatory agents, but have an inflammation diminishing function and are natural and free of side effects.

The invention provides biomarker profiles of cellular metabolites and methods for screening chemical compounds including pharmaceutical agents, lead and candidate drug compounds and other chemicals using human embryonic stem cells (hESC) or lineage-specific cells produced therefrom. The inventive methods are useful for testing toxicity, particularly developmental toxicity and detecting teratogenic effects of such chemical compounds.

The invention relates to a Lactobacillus gasseri strain CKCC 1913 and application thereof, the strain is preserved in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms on August 23, 2021, and the preservation number is CGMCC No.23175. The Lactobacillus gasseri strain CKCC 1913 has the advantages that the Lactobacillus gasseri strain CKCC 1913 can be used for preparing the lactobacillus gasseri strain CKCC 1913; a bacterial suspension, a cell metabolite and a cell content of the lactobacillus gasseri CKCC 1913 have a very good inhibition effect on the activity of dipeptidyl peptidase-4 and alpha-glucosidase, have antioxidant activity and can be used for preparing hypoglycemic drugs and antioxidant products. In addition, the strain CKCC 1913 has high gastrointestinal fluid tolerance and good gastric mucosa protein adhesion capacity, the viable count of CKCC 1913 single-strain fermented milk in the 21-day storage period is basically kept unchanged, and the acidity change is lower than 20 DEG T. Comprehensive research results show that the strain CKCC 1913 can be used as probiotics for reducing blood sugar and increasing the viable count of fermented milk products in the shelf life, and has huge application value.

To estimate concentration of a cellular metabolite contained in a culture medium where certain cells are cultured, using a simple method, etc., spectroscopy. The number of the certain cells is estimatable by applying previously obtained information regarding relationship between a cellular metabolite concentration and the number of the certain cells.In spectroscopy, the higher the cellular metabolite concentration, the more accurately the actual concentration of the cell consumed-substance can be estimated. Accordingly, the concentration of a cell-consumed substance, decreasing as cells are cultured, are estimated in the early to middle stages of culture, and the concentrations of a cell-produced substance, increasing as cells are cultured, are estimated in the middle to late stage of the culture. This enables estimation of the number of cells in the entire range from beginning to end of cell culture.

The invention provides a preparation method of a composite microecological preparation containing a fermentation medium. The composite microecological preparation is prepared by the following steps: yeast and L-valine-producing bacteria are subjected to combined fermentation culture in a solid medium containing betaine, and then subjected to bacteriolysis treatment and a self-chelating reaction. The composite microecological preparation contains cell metabolites such as cell contents, betaine and L-valine. The composite microecological preparation of the invention is rich in bacterial cell wall fragments, multivitamins, minerals, small peptides, organic acids, vitamins, nucleotides, oligosaccharides, polysaccharides and various growth-promoting factors, and the like, and can effectively improve the utilization rate of a feed to animals.

The invention provides a genetically modified micro-organism for intracellular biosynthesis of a cellular metabolite, comprising a synthetic error correction system having a penalty gene, whose expression leads to arrested growth or cell death (e.g. a toxin gene) in combination with a survival gene, whose expression provides an antidote that restores cell viability and normal growth (e.g. a cognate antitoxin gene). Alternatively, the system has a survival gene, alone, whose expression is essential for growth (i.e. essential gene). The synthetic error correction system further comprises a biosensor, whose function is to induce expression of the survival gene which leads to cell growth, only, when the cell produces a pre-defined level of a given metabolite. The invention further encompasses: a method for producing the genetically modified micro-organism; a method for producing a cellular metabolite with the genetically modified micro-organism; and use of the genetically modified micro-organism for producing a cellular metabolite.

The invention discloses a preparation method and application of a bone repair material having a multi-dimensional channel structure. The bone repair material is formed by stacking at least two laminated substrate structure layers, wherein each of the substrate structures is provided with through holes and grooves, and each of the substrate structures is prepared through substrate material preparation, substrate prefabricating, filler preparation, filling, stacking, forming, heat processing and heat treatment. According to the bone repair material prepared by the invention, three-dimensional network channels are constructed by the through holes and the grooves of the substrate structures and the channels are filled with collagen fibers, so that the bone repair material is conducive to cell adhesion growth and the in-growth of blood vessels and nerves, and the bone repair material is also convenient for transfer of nutrient substances and excretion of cell metabolites; meanwhile, cells are uniformly distributed inside the material, which is conducive to the formation of uniform and consistent tissues inside a stent; and the structural material can achieve firm biological chimerism with new bone tissues and can keep a good structural stability during degrading.

The invention discloses a preparing method of plant cell enzymolysis extracts for treating avian influenza. The method includes the following steps of weighing fresh garlic and bitter gourd to be cleaned, mixing and smashing the garlic and bitter gourd until the granularity of 10-20 mesh is reached, adding mature vinegar and edible grain liquor into an enzymolysis container, adding starch protease into the enzymolysis container to be rotated and stirred for 20 minutes at a high speed once every day, adding a chitosan oligosaccharide cell fusion enzyme after stirring is continuously conducted for 5 days, mixing a growth promoting agent in cooperation for standing for 30 days, and conducting clarifying, filtering, separating and extracting on fused cell metabolites at a normal temperature. According to the preparing method of the plant cell enzymolysis extracts for treating avian influenza, the garlic and bitter gourd are pure natural medicine and food homologous plant objects, substances extracted after cell fusion enzymolysis keep the existing characteristics of the garlic and bitter gourd of immunity, inflammation eliminating, bacterial killing, virus resisting and the like and also have the medicine property of killing avian influenza viruses, the side effects and medicine tolerance caused by the use of a large amount of antibiotics are avoided, and the survival rate of poultry is increased.