29 results about "Collagenase" patented technology

The present invention provides a composition comprising collagenase as an enzyme; calcium, histidine, and glycine as coenzymes, and a method for stabilizing enzymes, the method comprising the preparation of the composition by adding histidine and glycine to collagenase. According to the present invention, the enzyme composition and method for stabilizing enzymes can minimize or inhibit the agglutination of enzymes, maintain the concentration of freeze-dried enzymes even during long-term storage, and provide a formulation that has excellent rehydration time and hygroscopicity when restored to a liquid state, as well as superior enzyme activity.

The present application relates to a kind of anti-aging composition and its application of brightening and shrinking pores.The composition includes the following components: halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract, and the mass ratio of the halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract is (0.2-2):(0.001-0.1):(0.001-0.1):(0.001-0.1):(0.001-0.1).The composition of the present application can effectively improve the clearance rate of collagenase, maintain the elasticity and tightness of skin, have good anti-aging and pore-shrinking effect, and can also effectively remove hydroxyl radicals, have good whitening and brightening effect.

The present application relates to cell preparation process in the field of bioengineering, disclose a kind of high-efficiency preparation mouse primary hepatocyte method, comprising: perfusion needle is placed in reverse direction to liver direction through inferior vena cava, utilize the blood vessel clamp with elastic micro-tooth damping surface to lock the perfusion needle and inferior vena cava wall;The overflow damping of hepatic portal vein outflow end is dynamically regulated by blood vessel clamp, to construct controlled positive pressure permeation environment in liver vascular bed;Collagenase solution is radially permeated to liver parenchyma using the environment, the present application makes enzyme solution radial deflection and cross blood sinus barrier into di's cavity by constructing micro-pressure environment, eliminate the digestion dead zone originally due to flow path preference, ensure that enzyme molecule and extracellular matrix are uniformly contacted in global dimension, realize the collaborative promotion of tissue cutting efficiency and total amount of cell release, ensure that the output cell population has highly consistent physiological state.

This invention relates to the medical field, specifically to a cell digestion solution, a method for isolating γδT cells, and its application. The cell digestion solution comprises: collagenase, mammalian serum, antibiotics, a buffer, and a cell culture medium; wherein the collagenase content is 1-4 mg / ml, the mammalian serum content is 5 vol%-15 vol%, the antibiotic content is 0.5 vol%-2 vol%, and the buffer content is 5-15 mM / L. Through the synergistic effect and optimized content of components such as collagenase, mammalian serum, and buffer, the cell digestion solution effectively protects cell viability and surface antigen integrity while efficiently dissociating skin tissue, providing a high-quality cell source for subsequent isolation of rare immune cells.

This invention discloses a multi-stage separation and purification device for collagen peptides, relating to the field of collagen processing technology. It aims to solve the technical problem of difficulty in separating and purifying collagenase raw materials due to their diversity, as demonstrated by existing equipment. The device includes an enzymatic hydrolysis tank with a multi-stage separation mechanism at its bottom. This mechanism comprises a drive rod assembly, a fixed cylinder assembly, and a centrifugal inner ball assembly. The drive rod assembly drives the centrifugal inner ball assembly to rotate within the fixed cylinder assembly. The centrifugal inner ball assembly expands radially due to centrifugal force, forming a variable-gap extrusion and grinding chamber with the inner wall of the fixed cylinder assembly. It has two working states: bone fragment crushing and fiber chopping. The centrifugal ball segments have grooves, within which arc blades are movably inserted. These arc blades automatically extend and retract according to the rotational speed under the control of an elastic telescopic rod. A linked defoaming mechanism includes a guide plate and a defoaming disc, using rotational power to guide the foam to the vortex blades for rupture. This invention can simultaneously achieve bone fragment crushing, fiber depolymerization, and multi-stage filtration and defoaming, significantly improving raw material utilization and purification efficiency.

The application discloses an integrin-targeting peptide-collagenase drug combination and a drug composition constructed by the integrin-targeting peptide-collagenase drug combination. The drug combination can effectively block mechanical signals and biochemical signals mediating fibrosis, efficiently inhibit fibroblast activation, reduce the secretion of extracellular matrix proteins, and reshape the fibrotic microenvironment. Efficient reduction of fibrosis can promote the penetration of subsequent therapeutic drugs, and has a cascade penetration effect. The constructed drug composition can synergistically deliver integrin-targeting peptides and collagenase, and after release, the integrin-targeting peptides and collagenase can synergistically act in the form of free molecules in fibrotic lesions, thereby providing a new strategy for efficient delivery of fibrosis treatment drugs.

The application relates to a method for separating and primary culturing of monopterus albus liver cells. The method comprises the following steps: obtaining monopterus albus liver tissue, perfusing the liver tissue with HBSS buffer solution without calcium and magnesium until blood is completely drained; removing the capsule, blood clots and connective tissue after rinsing, and cutting the liver tissue; sequentially using 0.02% EDTA solution, 0.25% trypsin solution and 50 U / mL collagenase IV working solution to perform three-step digestion treatment on the liver tissue fragments; obtaining a cell suspension by blowing and filtering after the digestion is completed, and then performing gradient centrifugal purification of the cells at rotating speeds of 600 rpm, 250 rpm and 80 rpm in sequence; and finally resuspending the cells in DMEM / F12 culture medium containing 10% FBS and inoculating and culturing the cells. The monopterus albus primary liver cells obtained by the method have the advantages of large quantity, high activity and high purity, and provide a reliable in-vitro model for monopterus albus related research.

This invention belongs to the field of genetic engineering technology and discloses a high-efficiency expression method for recombinant collagenase and its application in the preparation of bovine bone collagen peptides. The collagenase gene derived from Bacillus sp. HUB-I-004 strain (accession number J9CTX3) was optimized using E. coli-preferred codons. A recombinant expression vector was constructed and transformed into E. coli BL21(DE3). After optimization of induction conditions and Ni-NTA affinity purification, a highly active recombinant collagenase with a specific activity of 231.05 U / mg was obtained. Bovine bone collagen was extracted using a high-temperature, high-pressure method. Bovine bone collagen peptides were prepared under optimal conditions of a substrate concentration of 10 mg / mL, an enzyme addition of 20%, and enzymatic hydrolysis at 37℃ for 8 h, significantly improving peptide yield. The obtained product is mainly composed of small molecule peptides and exhibits significant osteoblast proliferation-promoting activity, making it widely applicable in the preparation of health foods and pharmaceuticals.

The application discloses collagen tripeptide and a preparation method and application thereof, and belongs to the technical field of bioengineering. The preparation method comprises the following steps: obtaining bacillus velezensis with high collagenase yield through separation and identification, the bacillus velezensis belongs to probiotics, and high-activity collagenase is obtained through fermentation culture and purification; taking shark cartilage or blue fish skin as raw materials, and obtaining collagen tripeptide through the following steps of combined enzymolysis of alkaline protease and collagenase after pretreatment, inactivation, centrifugal separation and freeze-drying. The collagen tripeptide prepared by the application has a molecular weight of less than 500 Dalton, contains active components such as GHK tripeptide, is easy to absorb and has high safety, and can be used for preparing antioxidant products, anti-inflammatory products, memory-enhancing products or immunity-enhancing products. The application solves the problems of unsafe collagenase source and low collagen tripeptide preparation efficiency, and has a good industrial application prospect.

This invention relates to the field of cell engineering technology, and more particularly to a method for preparing immunomodulatory stem cells for treating graft-versus-host disease (GVHD). The invention provides a method for preparing immunomodulatory stem cells for treating GVHD, comprising the following steps: (1) washing human fat aspirate and digesting it with collagenase to obtain monocytes; (2) inoculating the monocytes obtained in step (1) into differentiation medium and culturing for 2–3 days, then continuing to culture the adherent cells for 18–22 days to obtain mesenchymal stem cells; (3) inoculating the mesenchymal stem cells obtained in step (2) into induction medium for induction culture, and after screening, obtaining immunomodulatory stem cells. The preparation method provided by this invention solves the technical problems of unstable sources, functional decline, and poor therapeutic effects of immunomodulatory stem cells in the prior art, providing a new, efficient, safe, and standardized solution for the treatment of GVHD.

This invention belongs to the field of stem cell sorting technology and discloses a method for preparing canine adipose-derived MUSE cells using magnetic beads and its application. The method first involves digesting canine adipose tissue with collagenase and then expanding it by adhesion. Subsequently, FITC-labeled SSEA-3 antibody is combined with anti-FITC magnetic beads, and SSEA-3 positive cells are captured using EasySep™ column-free magnetic bead sorting technology. The sorted positive cells are then seeded into a microsphere culture apparatus containing 81 microwells for induction culture. Experiments show that the purity of the MUSE cells obtained by this method is over 90%, and the spontaneously reconstructed colonies (M-clusters) in the microwell system exhibit strong SSEA-3 fluorescent expression. This invention effectively solves the problems of low initial content of canine MUSE cells and significant damage during flow cytometry sorting, providing a high-quality source of seed cells for canine regenerative medicine treatment.

Disclosed herein are methods of reducing collagenase-mediated bruising in a subject having cellulite, methods of treating cellulite in a subject, and collagenase-containing formulations.

Provided are a fermented apple composition having an effect of reducing or inhibiting UV-induced collagenase expression, and use thereof in the skin improvement and in the prevention or inhibition of skin photoaging.

Intracerebral hemorrhage (ICH), defined as spontaneous bleeding into the brain, is the deadliest, most disabling, and least treatable form of stroke. The aim of the present invention is to generate large amount of hemostatic mEVs to be used as hemostatic patches in different preclinical models of ICH. Indeed, the exogenous mEVs pf the present invention bearing TF and PSGL-1 improve outcome after collagenase-induced ICH by acting as intravascular hemostatic patches. The present invention thus relates to monocyte extracellular vesicles (mEVs) functionalized with tissue factor (TF) and P-Selectin Glycoprotein Ligand 1 (PSGL-1).

The application discloses a kind of Vibration Mountain beard toad hind leg / toe tissue cells and its quick acquisition method and application.The method comprises: the body surface disinfection of Vibration Mountain beard individual, cut and take hind leg or toe tissue, and put back after wound disinfection;After tissue is soaked in 75% alcohol and washed with PBS containing penicillin, streptomycin and amphotericin B, it is temporarily stored in 4 ℃ temporary storage solution;After tissue is cut, it is sequentially digested with trypsin and type I collagenase;In the optimized low permeability medium, primary culture is carried out at 25-27 ℃;When cell confluence reaches 60%-70%, it is passaged using trypsin-EDTA digestion;Freezing and recovery.The application realizes the non-lethal cell of Vibration Mountain beard for the first time Quick acquisition, cell first time migration time is only 3 days, grow into monolayer only 12-17 days, pollution rate is reduced to 5.6%, recovery survival rate reaches 76.19%, and provides efficient and reliable technical support for endangered amphibian germplasm resource preservation and biological research.

The application discloses a preparation method and application of a recombinant collagenase type I and a mutant thereof. A mutant of the collagenase type I is provided in the application, and the mutant has good alkali resistance and can be applied to enzymatic extraction of fish skin waste. The application further provides an expression method of the recombinant collagenase type I and the mutant thereof based on a prokaryotic expression system. The expression method can improve the expression amount of soluble protein, and the product has high activity and is suitable for industrial amplification production.

The present application belongs to the field of microbial technology, and more specifically, the present application relates to an inactivated culture of plant lactobacillus BGI-N6, its use and products containing the inactivated culture. The present application provides an inactivated culture product of probiotics, which can break through the condition limitation of application of active probiotics, effectively eliminate free radicals, inhibit collagenase and elastase, inhibit skin pathogenic bacteria, so as to achieve better effect of slowing down skin aging.

This application provides a method for preparing collagen peptides for sports nutrition foods used in bone and joint recovery. The method includes the following steps: S1: dispersing collagen raw materials in water to obtain a collagen dispersion; S2: adding an amphoteric molecule and a soluble divalent metal salt to the collagen dispersion to obtain a pretreated dispersion; S3: adding an endopeptide to the pretreated dispersion to obtain an enzymatic hydrolysate; S4: post-processing and drying the enzymatic hydrolysate to obtain collagen peptides. This application, by regulating the collagen dispersion system before enzymatic hydrolysis and continuously adjusting the dispersion state of the peptide chains during enzymatic hydrolysis, can reduce the aggregation tendency between collagen peptide chains, resulting in collagen peptides with better dissolution rate and solution stability, thus making them more suitable for use in sports nutrition foods for bone and joint recovery.

This invention belongs to the field of cosmetic technology, specifically relating to an anti-aging composition containing nanospheres and its preparation method, as well as its application in skincare products. The method employs a double emulsion solvent evaporation method to prepare nanospheres. Soybean lecithin and polylactic acid-glycolic acid copolymer are dissolved in an organic solvent as the oil phase. Superoxide dismutase, collagenase, trehalose, and bovine serum albumin are dissolved in phosphate buffer as the inner aqueous phase. After emulsification to form a primary emulsion, it is added to an outer aqueous phase containing polyvinyl alcohol and homogenized to form a double emulsion. After solidification, centrifugation, and washing, the nanospheres are obtained. The resulting composition exhibits good enzyme protection and skin permeability, effectively scavenging free radicals and inhibiting collagen degradation, showing broad application prospects in the skincare field. This invention features a simple process, uses commercially available raw materials, and is suitable for large-scale production.

This invention relates to the field of animal model construction technology, and more particularly to a method for constructing a zebrafish coronary artery microcirculation injury model based on enzyme stripping and its application. This invention, for the first time, utilizes a rationally composed mixture of collagenases to specifically strip coronary artery endothelial cells, minimizing non-specific damage to cardiomyocytes and stably inducing coronary artery injury to construct a zebrafish model of coronary artery injury, particularly coronary artery microcirculation injury. This model not only has a highly localized injury area and minimal impact on the myocardium, but also exhibits a low mortality rate. Furthermore, the zebrafish model of coronary artery injury constructed by this invention demonstrates significant regenerative potential, with its blood vessels exhibiting strong regenerative capacity. It also shows high sensitivity and responsiveness to drugs, making it potentially valuable in drug screening and mechanism research.

This invention provides a method for isolating spermatogonial stem cells from frozen testicular tissue, belonging to the field of cell biology. The isolation method provided by this invention includes the following steps: thawing frozen testicular tissue; digesting the thawed tissue with collagenase IV and DNase I until the seminiferous tubules are exposed and their walls become rough; washing with DPBS buffer, centrifuging, and culturing until cells migrate; collecting the migrated cells and purifying them for spermatogonial stem cell enrichment. The method described in this invention can maximally preserve the internal environment of spermatogonial stem cells, maintain cell viability, and reduce cell damage, and can be used for isolating spermatogonial stem cells from frozen testicular tissue.

The utility model provides a kind of fish collagenase hydrolysate is carried out rough filtration's separation device, it includes frame, motor, driving rod, first gear, second gear, lead screw, lifting seat, sliding block, slide rail, filter box, by motor, gear meshing and lead screw thread pair linkage, it is converted into lifting seat vertical lifting with horizontal rotation, filter box is lifted and moves stably, avoid enzymatic solution spillage;Slide rail orientation ensures accurate positioning, and replaceable filter core plate realizes continuous feeding, automatic lifting replaces manual handling, improves rough filtration efficiency and safety, overall device is automatically lifted by mechanical linkage, replaces manual handling, reduces labor intensity, improves operating safety.

Products, processes, compositions, kits, and methods are provided for cartilage-derived implants. The implants can exhibit resistance to enzyme (e.g., collagenase, protease, etc.) digestion compared to the source tissue from which they were derived while still having one or more mechanical properties comparable to the source tissue from which they were derived. The implants can also have a plurality of molecular bridges between molecules of the cartilaginous material. The molecular bridges can connect one or more collagen fibrils and / or / with one or more glycosaminoglycans. The implants can also be treated with cationic detergent, packaged and sterilized with or without additional components, and surgically implanted into subjects.

This invention provides a method for isolating and identifying interstitial cells from Bactrian camels, belonging to the field of cell isolation technology. A mixture of 0.08–0.12% collagenase IV solution and 0.23–0.27% trypsin-EDTA at a volume ratio of 1:2 is used to digest Bactrian camel testicular tissue blocks. The mixture is then purified by hypotonic treatment to remove excess germ cells, differential adhesion purification to remove a large number of supporting cells, and Percoll density gradient centrifugation to remove a small number of supporting cells. The suspension, obtained from a 34%–60% Percoll gradient, is transferred to a new culture flask to obtain interstitial cells with a purity ≥95%. Identification is performed using morphological observation, immunofluorescence staining, and secretory function testing. The interstitial cells isolated from Bactrian camels using this invention can be used for research on the reproductive performance of Bactrian camels, laying the foundation for understanding the reproductive performance of Bactrian camels.

This invention discloses a method for detecting bacterial endotoxins in gelatin microcarriers used for cell culture, comprising the following steps: (1) rechecking the sensitivity of the Limulus Amebocyte Lysate (LAL) reagent to ensure that the sensitivity result of the LAL reagent is between 0.5 and 2.0 λ; (2) determining the limit value and minimum dilution concentration of bacterial endotoxins in the gelatin microcarriers; (3) enzymatically digesting the PBS solution of the gelatin microcarriers to be tested with collagenase solution to obtain an enzymatic digest; (4) checking for bacterial endotoxins in the gelatin microcarriers: mixing a magnesium ion regulator and the LAL reagent to obtain a reconstituted LAL reagent, mixing the enzymatic digest obtained in step (3) with the reconstituted LAL reagent to obtain a mixed solution, and using the gelation method to determine whether bacterial endotoxins are present. By combining the pretreatment enzymatic digestion step with the LAL reagent reconstituted with a magnesium ion regulator, the gelatin microcarriers can be effectively enzymatically digested, fully releasing endotoxins, and the interference of phosphate ions can be effectively eliminated by the magnesium ion regulator, thereby improving the specificity and accuracy of the detection.

Formulations, their fabrication methods, and methods of treating fibrosis include tunable hydrogels loaded with collagenase for breaking down collagen and thereby reducing stiffness in fibrotic tissue. The formulations may be tuned for topical application or injection, and provide localized and sustained treatment of fibrotic conditions.

This invention provides a crocodile bone collagen peptide powder for improving bone density, its preparation method, and its application, relating to the field of biopharmaceutical technology. The preparation method includes the following steps: crocodile bone is pulverized and mixed with sodium chloride solution, followed by a first separation to obtain bone residue; water is added and mixed, followed by a second separation to obtain defatted bone residue; the defatted bone residue is mixed with water and a deodorizing agent, stirred, and separated to obtain deodorized bone residue; the deodorized bone residue is added to water, and ultrasonic-assisted extraction is performed, separating and collecting the supernatant; the supernatant is mixed with collagenase and trypsin for a first enzymatic hydrolysis; carboxypeptidase A and proline endopeptidase are added for a second enzymatic hydrolysis; the supernatant is separated, and transglutaminase is added to react, separating and obtaining the enzymatic hydrolysate; the enzymatic hydrolysate is separated using a multi-stage tangential flow membrane separation system, collecting the retentate from a 500 Da nanofiltration membrane; the final product is concentrated and dried. The crocodile bone collagen peptide powder obtained by this invention through a specific preparation method significantly improves taste, enhances stability, and strengthens the effect of improving bone density and bone calcium.