Method for producing viable breast cells |
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IPC classes for russian patent Method for producing viable breast cells (RU 2409664):
 Method for producing dedifferentiated cells of adult retinal pigment eye epithelium / 2409663
Eyeball is enucleated in autopsy, washed in an alcoholic solution and in Ca2+ Mg2+ -free Hanks' solution with an antibiotic added; an anterior segment of an eyeball is removed along a dentate line, a vitreous body and a neutral retina are separated from a pigment epithelium; an eyecup is filled with Ca2+ Mg2+ -free Hanks' solution with EDTA, and retinal pigment epithelial cells are incubated for 15-30 min; the produced cells are pipetted and transferred to a sterile recovery medium, pipetted, centrifuged; the recovered cells are resuspended in a high-serum growth medium, then the prepared suspension of the retinal pigment epithelial cells is distributed over a culture surfaces and cultivated until the attached cells reach 15-25 % of its area; the suspension with the unattached cells is aspirated, distributed over the fresh culture surfaces and cultivated; and the unattached cells are added with the high-serum growth medium and cultivated until a confluent finish monolayer is produced.
 Method for producing donor chondrocytes / 2409662
Donor chondrocytes are produced by recovering and grinding donor cartilaginous tissue, carrying out enzymatic treatment, filtration and centrifugation of the prepared chondrocyte suspension and cultivation thereof in a nutrient medium DMEM with added 20 % FBS and perphthorane in amount 15-25 % of total amount of the nutrient medium.
Method for producing donor chondrocytes / 2409662
Donor chondrocytes are produced by recovering and grinding donor cartilaginous tissue, carrying out enzymatic treatment, filtration and centrifugation of the prepared chondrocyte suspension and cultivation thereof in a nutrient medium DMEM with added 20 % FBS and perphthorane in amount 15-25 % of total amount of the nutrient medium.
 Compositions and methods for labelling biological compounds with stable isotope / 2409657
Microorganisms are used to prepare a culture medium, where the microorganisms are cultured on a mineral medium which supports growth of that organism, where essentially all assimilated atoms are isotope-labelled to obtain a labelled biomass. The obtained biomass undergoes autolysis to obtain a labelled autolysate. This biomass can be further extracted with an organic solvent to obtain lipids. The delipidised biomass is hydrolysed to obtain labelled amino acids and other nutrient substances which are used together with the autolysate extracted by lipids and additional components to prepare a culture medium for growing mammal or insect cells in the culture for synthesis of biomolecules. The obtained biomolecule is a mammal membrane protein, particularly in which 20-100% hydrogen atoms in that protein are totally substituted with a 2H isotope.
 Version of group i poaceae allergen characterised by lower allergenicity and maintained t-cell responsiveness (versions), coding dna molecule and application thereof / 2409589
Invention refers to producing versions of group I Poaceae (holy grass) allergen, also can be used either for specific immunotherapy (hyposensitisation) of patients with grass pollen allergy, or for preventive immunotherapy of grass pollen allergies. The produced versions are characterised by Cys41 Ser, Cys57Ser, Cys69Ser, Cys72Ser, Cys77Ser, Cys83Ser and Cysl39Ser substitutes in a Phi p1 mature protein sequence. Also, a structure of the allergen versions can be presented with no fragments relevant to amino acid residues 1-6, 1-30, 92-104, 115-119, 175-185 and 213-220 or 1-6, 115-119 and 213-220 as a part of a primary sequence of Phi p1 mature protein.
 Method of neuronal cell differentiation from embryo stem (es) / 2407790
Invention refers to biotechnology, particularly to a method of induced differentiation of embryo stem (ES) cells in neuronal precursor cells. The presented method involves ES cell culture by sowing the ES cells of density approximately 0.5 × 105 - 2 × 105 cells in cm2 and dissociating the ES cells 2 days after sowing. Then, cell aggregates (CA) are formed that involves sampling the cells of high proliferative activity of doubling time within 0 to 24 hours and sowing these cells of density approximately 0.5 × 105 - 5 × 105 cells in ml to form the CAs. Further, the cell aggregates are processed with retinoic acid (RA). Then, the CAs are dissociated to prepare a neuronal precursor cell culture.
Method of neuronal cell differentiation from embryo stem (es) / 2407790
Invention refers to biotechnology, particularly to a method of induced differentiation of embryo stem (ES) cells in neuronal precursor cells. The presented method involves ES cell culture by sowing the ES cells of density approximately 0.5 × 105 - 2 × 105 cells in cm2 and dissociating the ES cells 2 days after sowing. Then, cell aggregates (CA) are formed that involves sampling the cells of high proliferative activity of doubling time within 0 to 24 hours and sowing these cells of density approximately 0.5 × 105 - 5 × 105 cells in ml to form the CAs. Further, the cell aggregates are processed with retinoic acid (RA). Then, the CAs are dissociated to prepare a neuronal precursor cell culture.
Method of neuronal cell differentiation from embryo stem (es) / 2407790
Invention refers to biotechnology, particularly to a method of induced differentiation of embryo stem (ES) cells in neuronal precursor cells. The presented method involves ES cell culture by sowing the ES cells of density approximately 0.5 × 105 - 2 × 105 cells in cm2 and dissociating the ES cells 2 days after sowing. Then, cell aggregates (CA) are formed that involves sampling the cells of high proliferative activity of doubling time within 0 to 24 hours and sowing these cells of density approximately 0.5 × 105 - 5 × 105 cells in ml to form the CAs. Further, the cell aggregates are processed with retinoic acid (RA). Then, the CAs are dissociated to prepare a neuronal precursor cell culture.
 Method for producing live cell vaccine for breast cancer prevention / 2407789
Murine benign spontaneous breast tumour cells are cultivated. At first, the cultivation procedure is in vitro in the DMEM/RPMI-1640 medium with 10 % of bovine embryo serum up to 20 passages. Then, in vivo up to 22 passages. Then, once again in vitro up to 25 passages. Then, 10 thousand-1 million produced cells are introduced in a connective tissue capsule formed in the Balb/c mice after the introduction of a biocompatible gel. The tumour growth is enabled. The encapsulated tumour is aseptically removed and trypsinised to produce primary epithelial cell cultures. The produced cells concentrated 10 thousand-1 million are introduced into the capsule subcutaneously gel-formed in the Balb/c mice. The tumour growth is enabled for vaccination. In the in vitro culture, the cells are transplanted once a week while being disseminated in the ratio 1:3 - 1:4, with a monolayer twice washed with EDTA solution and removed with 0.25% trypsin during 2-3 min at room temperature, and the trypsinisation process is terminated by adding 10 % of fetal serum "к.р.с.".
 Agent for increasing reserved stem cell number in organism / 2405822
Invention refers to radiation synthesis nanotechnology for the creation of a low-immunogenicity agent to increase the reserved stem cell number in an organism, and can be used in regenerative medicine. The agent represents polymer immobilised hyaluronidase introduced in the dissolved low-molecular water-soluble polymer pre-exposed to ionising radiation in dosage 1-5 Mrad, to final concentration 70 UN in 1 ml.
 Novel cross-linking reagents for producing chitosan-based biocompatible materials / 2408618
Articles based on biologically active polymer materials can be used in surgery when treating wounds and as materials for temporary replacement of body tissue, in biotechnology for obtaining matrices for growing cell cultures, in pharmaceuticals as carriers of enzymes and other biologically active compounds. The cross-linking reagents are 2,4-derivatives of 3-oxa glutaric dialdehyde (2,2'-oxydiacetaldehyde). These compounds can be obtained via periodate oxidation of monosaccharides, nucleosides and nucleotides.
 Method of obtaining polysaccharide material / 2392972
Invention relates to a method of obtaining polysaccharide fibre for making materials, specifically for making surgical suture materials absorbable in a human and a mammal body, absorbable and non-absorbable dressing material and absorbable textile matrix materials. The method is characterised by that 2.4-4.0 wt % solution of polysaccharide in dimethylacetamide which contains 4.56-10.00 wt % lithium chloride is mixed with 1.0-5.0 wt % poly-N-vinylpyrrolidone with molecular weight of 8-35 kDa or a metallopolymer complex - fine-grained silver stabilised by poly-N-vinylpyrrolidone in such an amount that, content of fine-grained silver relative polysaccharide dissolved in spinning solution ranges from 0.07 to 0.87 wt %. Mass ratio of polysaccharide: metallopolymer complex equals 88.0-99.0:1.0-12 wt %. The mixture is intensely stirred, held, filtered, degassed and the obtained spinning solution is extruded at room temperature into an alcohol deposition tank in form of water-soluble aliphatic C2 and C3 alcohols. The fibre is then processed in plastification and washing tanks and dried.
 Taxanes covalently bonded with hyaluronic acid or hyaluronic acid derivatives / 2384593
Invention relates to taxane, especially to paclitaxel and docetaxel covalently bonded with hyaluronic acid or a hyaluronic acid derivative, used as active substance in pharmaceutical compositions which are used in oncology, for treating tumours, autoimmune diseases and restenosis, as well as a coating for stents and medical devices. The covalent bond is formed between hydroxyl groups of taxane and carboxyl groups or hydroxyl groups of hyaluronic acid or hyaluronic acid derivatives, or amino groups of deacetylated hyaluronic acid. Bonding can take place using a linking compound (spacer compound) which bonds taxane with hyaluronic acid or hyaluronic acid derivative under the condition that, the linking compound is different from hydrazide.
Composition - osteoinducer and osteoconductor in treatment of bone pathology in dentistry and oral surgery and method of obtaining composition - osteoinducer and osteoconductor in treatment of bone pathology in dentistry and oral surgery / 2383564
Composition includes chitosan gel, which has bactericidal properties, representing matrix for including in it of water solution for "Adgelon" injection, which contains serum glycoprotein from blood of livestock, possessing biological activity in supersmall doses 10-9-10-15 mg/ml, and calcium salt - preparation "Adgelon". In obtaining composition medicine "Adgelon" is immobilised on chitosan gel.
 Complex matrix for medico-biological application / 2360928
Complex matrix consists of at least one biologically compatible polymer of natural origin, structured with sewing agent, which represents two- or multi-functional molecule, selected from epoxides, epihalohydrines and divinyl sulphone, on said polymer inoculated are chains with molecular weight less than 50000 daltons, selected from polymers of natural origin of small size, preferably, derivatives of cellulose or other biological polymer derivatives which naturally are not present in human organism, and/or non-polymerised chains with properties of oxidation inhibitors or ability to inhibit reactions of matrix decomposition, preferably, vitamins, enzymes or molecules, consisting of one or several cycles, degree of inoculation, expressed as ratio of moles of inoculated molecules and quantity of moles of polymer units, constitutes from 10 to 40%. Also described are method of obtaining such matrix and its application for separation, replacement, filling or addition of biological fluid or tissues.
Method for production of modified paper / 2355842
Invention relates to papermaking technology, precisely to production of modified paper with higher gas-proof and heat-protective properties, and can be applied in constructions, aircraft and automobile constructions, shipbuilding. The method includes treatment of paper with the mixture of 5-7% aqueous solution of polyvinyl alcohol with 5-7% aqueous solution of chitosan at their ratio 1:1 within 10-15 minutes, thereafter treatment with 15-20% aqueous solution of methyl phosphate borate and drying.
Compound for production of modified paper / 2355841
Invention relates to production technology of synthetic paper, precisely to production of modified paper with higher gas-proof and heat-protective properties, and can be applied in constructions, aircraft and automobile constructions, shipbuilding. The compound contains 5-7% aqueous solution of polyvinyl alcohol, 15-20% aqueous solution of methyl phosphate borate and 5-7% aqueous solution of chitosan at the following ratio, pts. wt. polyvinyl alcohol - 5-7; chitosan - 5-7; methyl phosphate borate - 15-20; water - 275-266.
 Chitosanium perchlorate, method for its preparing and power-consuming composition comprising thereof / 2315774
Invention describes chitosanium perchlorate of the following formula: C6O4H9NH3ClO4. This compound shows such properties as explosion-proof, absence of toxicity, resistance to moisture and heating, mechanical effects and possesses high oxidizing and binding properties. This allows its using in power-consuming compositions in mixture with chitosanium dodecahydro-clozododecaborate wherein it functions as a binding agent. The quantitative ratio between chitosanium dodecahydro-clozododecaborate in the composition is determined by required regimen of combustion: the more content of chitosanium perchlorate the higher activity of the composition.
 Chitosan product, method for its preparing (variants) / 2313538
Invention relates to preparing biologically active chitosan substances and their derivatives. Invention describes a modified chitosan substance showing pH-neutral reaction and plastic structure of chitosan particles as fractal chitosan particles of size of nanofractals from 1 nm, not less, and to 5000 nm, not above, or as cross-linked net-shaped polymer having multiple cavities of size from 1 nm, not less, to 50 nm, not above. Invention describes methods for their preparing. Invention provides high transdermal penetration of chitosan substance and enhanced capacity for administration of medicinal or biologically active substances into chitosan substance. Invention can be used in manufacturing cosmetic, curative-cosmetic, pharmacological preparations, biologically active food supplements and foodstuffs.
Chitosan-base composition reducing nitrate content in plant growing production / 2257711
Invention describes a chitosan-base composition reducing the content of nitrates in plant growing production. The composition comprises chitosan of molecular mass 20-150 kDa, organic acids - mixture of succinic, ascorbic and sorbic acids in the mass ratio = 2:1:1, respectively, activating agents of nitrate and nitrite reductases - iron nitrate, ammonium molybdate, indolyl acetic acid and ethylenediaminetetraacetic acid in the mass ratio = 20:5:1:1, respectively; or iron nitrate, ammonium molybdate, indolyl acetic acid and N,N-dicarboxymethylglutamic acid in the mass ratio = 20:5:1:1, respectively, and substances enhancing synthesis of chlorophyll - 2-oxoglutaric acid and L-glutamic acid in the mass ratio = 30:1, and a surface-active substance also in the following ratio of above indicated components, wt.-%: chitosan, 15-25; organic acids, 15-25; activators of nitrate and nitrite reductases, 25-30; enhancers of chlorophyll synthesis, 29-35, and surface-active substance, 1-3. The composition provides enhancing assimilation of inorganic nitrogen in plant and reduces the content of nitrates in plant growing productions.
 Nanocomposite and method of its production / 2256601
Proposed nanocomposite can be used as component contributing to charges of consumer properties of materials made on its base. Nanocomposite includes fibrils of filler-chitin individualized to nanosizes with distance between fibrils from 709 to 20-22 nm and water-soluble polymeric matrix in interfibril space. Degree of filling of nanocomposite is 0.05-0.25% mass. Fibrils are arranged in parallel and they have cross size of 4 nm. Method of production of nanocomposite comes to the following: free-radical polymerization in water medium of at least one monomer of row of acrylic acid, salt of acrylic acid, acrylamide is carried out in presence of filler. Initiator is chosen from the row of water-soluble peroxides, hydroperoxides or their salts, potassium persulfate. Individualization to nanosizes of fibrils is done simultaneously with process of polymerization and/or with combination of said process with mechanical disintegrating action by disintegrating or pressing, or pressing with abrasion shift. Nanocomposite is obtained in form of film, being pervaporation membrane.
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FIELD: medicine. SUBSTANCE: solution of viable breast cells after enzymatic degradation in a collagenase solution of analysed tissue at temperature 37°C for 30-35 minutes, are applied on a biomaterial of a native form of hyaluronic acid, stored at room temperature and constant humidity 30-50 %, and cell viability is controlled by discoloration of cell monolayer. EFFECT: invention allows producing a viable cell monolayer and providing their vital activity for 2-3 hours. 1 tbl, 2 dwg
The invention relates to biotechnology and can be used in Cytology, histology, transplantation, Microbiology, medical research. The known method dissociation of cells developed on the example of hepatocytes (Application for invention No. 2005104557/13 dated 17.07.2003, BI No. 25, 2005). The method consists in the following. The analyzed tissue is placed in a solution of phosphate buffer with collagenase and Pancreatin. Aged 30-35 minutes at 37°C. followed by several cycles consisting of centrifugation, filtration. In the final stage, the solution of cells placed in culture medium to ensure their livelihoods. This method has significant drawbacks. Cells in the culture medium can be distributed not only on the surface, but throughout the thickness of the layer of culture medium, which adversely affects the measurement. After measurements of the cells were killed, and for further measurements of these cells must be cultivated. Multiple cycles consisting of filtration and centrifugation, significantly complicates the process. The technical result of the method is obtaining a monolayer of viable cells, and ensuring their livelihoods for 2-3 hours. The problem is solved in that in the method of obtaining viable glue is OK breast cancer, providing enzymatic cleavage in a solution of collagenase examined tissue at 37°C for 30-35 minutes, a solution of cells is applied to the biomaterial of hyaluronic acid, stored at room temperature and a constant humidity of 30-50%, and the monitoring of cell viability carry out the staining of monolayer cells. The figure 1 shows the distribution of cells on the biomaterial surface, showing the provision of biomaterial from hyaluronic acid, a unique adhesion imparted on the surface of cells. The application of cell suspension onto the surface of the biopolymer cells evenly distributed over its surface, while maintaining viability. The figure 2 shows the distribution of cells in culture medium. In contrast to the distribution on the biomaterial most cells in culture medium were distributed throughout its thickness. The method is as follows. As a source of cellular material for research selected normal and tumor cells of the mammary gland of mice BYRB, because the data of laboratory animals are considered to be the reference for such studies. Pieces of tumor and normal researched the breast tissue of mice BYRB placed in a tube with a solution of collagenase in phosphate buffer (pH 7.4) with conc what Tracia 0.5 mg/ml This solution with a piece of the investigated tissue incubated at 37°C with constant stirring. The optimal time for dissociation of tumor and healthy tissue is 30-35 minutes. After carrying out the enzymatic dissociation of cell suspension is applied to the biomaterial of hyaluronic acid (thickness of 0.25 mm, a density of 340.13 kg/m3)is a polymer of hyaluronic acid native form. This material meets the main criteria of biologically compatible matrix: the absence of cytotoxicity, maintaining adhesion, as shown in figure 1; fixing, proliferation and differentiation is placed on the surface of cells, the absence of an inflammatory response to the material and the immune response, a sufficient mechanical strength in accordance with the purpose, bioresorbability normal metabolic pathways (Agrawal CM et al. Biodegradable PLA/PGA polymers for tissue engineering in orthopaedica // Material Science Forum. - 1997. - P.115-128; KJL Burg et al. Biomateriats development for bone tissue engineering // Biomateriats. - 2000. No. 21. - R. 2347-2359). The biomaterial with cells stored at room temperature in a desiccator, in which immediately before placing the cells were filled with saline (0.9% NaCl)to maintain the biomaterial in the wet state. Immediately after applying the solution of the cells on the biomaterial cells are evenly distributed over the surface, forming a monolayer glue is OK. Reduce liquid in a drop of solution cells due to the evaporation of metabolic processes in cells and the absorption of moisture by the biomaterial is compensated by the addition of a phosphate buffer. Monitoring of cell viability carried out visually by color colors monolayer. In the case of living cells, the monolayer has a white color, in the case of dead cells - light brown. The choice of the biomaterial of hyaluronic acid due to the biological function of hyaluronic acid in naivnyh tissues (connective, epithelial tissue). Found that hyaluronic acid due to the unique physical qualities provides starecheski distribution of cells in the intercellular matrix and creates optimal conditions for their migration and mitosis (Brun P., Cortivo R, Radicc M, Abatangeio G. Hyaiuronan-based biomaterials in tissue engineering. New Frontiers in Medical Sciences: Redefining Hyaluronan // Symposium Proceedings, Padua, Italy. - June 1999. - P.269). The method was carried out at different humidity storage biomaterial coated with cells. The dependence of the biomaterial from humidity, which affects the viability of the cells included in the table.
The choice of wet conditions dictated by the hydrophilic properties of the biomaterial. When humidity less than 30% of the biomaterial structure does not allow the cells to optimally adhesives on its surface, resulting in viable cells within 10-15 minutes. When humidity is more than 50% disrupted the structure of the biomaterial, it becomes gel-like and unsuitable for layers of cells on its surface. Thus, the biomaterial is obtained monolayer cells and provided their jiznesposobnost is 2-3 hours. The method of obtaining viable cells of the breast, providing enzymatic cleavage in a solution of collagenase examined tissue at 37°C for 30-35 min, characterized in that after enzymatic cleavage solution of cells is applied to the biomaterial-based polymer native form of hyaluronic acid, stored at room temperature and a constant humidity of 30-50%, and the monitoring of cell viability carried out by a colour change of the monolayer of cells.
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