Method of separating leukocyte concentrate, which contains hemopoetic stem cells, from umbilical cord blood

FIELD: medicine.

SUBSTANCE: method related to field of medicine, namely to transfusiology. Method includes collecting umbilical cord blood (UCB) into container, weighing it with calculation of its volume, determining type of human leukocyte antigen for comparing hypocompatibility, presence or absence of infection, introduction into packet with UCB, anticoagulant and hydroxyethyl starch, mechanical mixing of packet content in various planes, incubation after mixing at room temperature with separation of erythrocyte mass (EM) by sedimentation, placement of centrifuge chamber in separator, connecting to it on symmetry axis main pipeline, the latter being connected via distribution unit via pipelines to sacks for collection of blood components and to packet with mixture of UCB, anticoagulant and hydroxyethyl starch. After that, chamber is filled with packet content, and mixture is subjected to separation, with its separation into EM, plasma and leukocyte concentrate with their distribution into corresponding sacks and into empty packet. After separation, viability and amount of SC are determined, they are checked on biological inoculation of sample of plasma and EM mixture, dose of plasma is tested for detection of antibodies, hemotransmissive infections, dose of EM is used to determine blood group and Rhesus factor.

EFFECT: method application makes it possible to increase quality and reliability of determining SM from various biological contaminations.

1 dwg

 

The method of separation of umbilical cord blood leukocyte concentrate containing hematopoietic stem cells relates to the field of medicine to the transfusion.

Cell therapy is a modern method of treatment of various diseases. Clinical application of innovative cell technologies aimed at improving the quality of life and increase life expectancy of the population. The use of new cell technologies in health permitted by Roszdravnadzor license No. 99-1-001458 from 12.05.2005, Stem cells are versatile precursor cells of all organs and blood cells. They are able to share under certain conditions and turn into any kind of tissue. This property is used when necessary, for example, replacement and restoration of functions of hematopoietic tissue. There are two main types of stem cells: hematopoietic and mesenchymal. Hematopoietic stem cells (HSC) are transformed into Mature blood cells, carry oxygen and nutrients to the tissues and are responsible for immunity and immunity to various diseases. Mesenchymal stem cells give rise to all the "specialized" cells and organs of the human body. Currently, there are methods of obtaining stem cells from bone MH is a, peripheral blood, umbilical cord blood, adipose tissue, dental pulp. Throughout the life of the person ages and is exposed to the adverse effects of the external environment and ecology. Factors such as radiation, cancerogene substances that affect the entire body and all its cells. Stem cells derived from umbilical cord blood, much younger than similar cells from the bone marrow, since they saved early in life and is more protected from the harmful effects of the number of cell divisions is limited. Stem cells from umbilical cord blood collected at one of the early stages of a person's life, so the ability of these cells to divide and turn into the desired body cells and their proliferative ability is higher. Umbilical cord blood is collected quickly and easily. By itself, the process of collecting cord blood is simple, takes less than 10 minutes, does not cause discomfort and safe for both mother and baby, because with them there is no contact.

Cord blood is collected by the midwife after clipping the umbilical cord can be collected after an independent childbirth, and after caesarean section. When a child is born, the midwife pinched and cut the umbilical cord, then give her the needle from the system, where blood flows. The collected blood was placed in a container for transportation. Blood is protected from shock, vibration, rez is their temperature and in complete safety along with the accompanying documentation is delivered to the store. Delivery is made within 24 hours, which allows for the collection of blood in almost any city of the Russian Federation. Before placing in the storage of umbilical cord blood of the baby should be treated. Blood weigh, count the number of stem cells. Then the stem cells extracted from umbilical cord blood and add to them cryoprotector - solution, which protects cells from destruction when thawed. Selected stem cells are placed in bags designed specifically for long-term storage at low temperatures, and three cryoprobes - "satellite". Each sample is assigned a unique identification number. Used marking guarantees the inviolability of the stem cells of the child. Stem cells will be stored in sealed sterile and airtight bag in the conditions of deep cold, see "Clinical Hematology. The Handbook. Companion of the doctor" edited Abdulkadyrov K.M., SPb., "PETER", 2006, str-398.

The disadvantages of the above method of separation of hematopoietic stem cells from umbilical cord blood are: the method is technologically inefficient, there are no steps to determine the quality of selected umbilical cord blood stem cells, there are no checks on the identification of potential or lack of biological contamination and infection rates, no what are the steps and quantitative values, allowing the process of obtaining hematopoietic stem cells.

In addition, a known method of isolation of hematopoietic stem cells from umbilical cord blood, see http://www.gemabank, EN/pub/nl.htmal-cma, authors: Camanducaia, Naamanka, Tab, Avellar "Actual problems of Hematology and blood transfusion. Harvesting and storage of umbilical cord blood. Materials of scientific-practical conference, St. Petersburg, 2000, the cord blood Collection is carried out at physiological childbirth after giving birth and separation from the placenta. After that make a puncture of a vein of the umbilical cord. The procedure of collecting cord blood lasts no more than 10 minutes. After blood collection container with umbilical cord blood is weighed to determine the mass of the collected blood. The recalculation of the mass of blood volume is performed by dividing the weight by the density of umbilical cord blood with the anticoagulant solution (gleyzer" - solution hydronitrate sodium), which is 104-105 g in 100 ml. Next in blood to determine the content of leukocytes, erythrocytes, platelets, hemoglobin, and also carry out bacteriological culture of sterility. Selection of red and white blood cells carry through sedimentatio in a 10% solution of hydroxyethylamine in correlation with cord blood 1:4, laundering from hydroxyethylstarch the Ala and separation with a speed of 1500 rpm for 10 minutes. Getting nucleated cells is carried out in a density gradient ficoll through separation and solution of hydroxyethylamine.

The disadvantages of such a method of selection of hematopoietic stem cells from umbilical cord blood are: insufficient quality, quantity and reliability of tests of umbilical cord blood, since there is no test for the presence of HLA - typing histocompatibility cord blood, there is no test for detecting antibodies T.pallidum in plasma, cord blood, no control actions and validation of plasma cord blood and there is no validation on bloodborne infection; the resulting fraction of nucleated cells in a density gradient ficoll can harm human health, the ratio of sedimentologica tools - hydroxyethylamino 1:4 is not effective, the supernatant pipetted, an open system can lead to infection, technology way identify hematopoietic stem cells practically and industrially developed and inefficient.

The technical result of the proposed method of selection of hematopoietic cells from umbilical cord blood is: improving the quality and reliability from different infectivity and biological contaminants collected and isolated from umbilical cord blood stem cells contained in the Le is azithrom concentrate, the improving technology of obtaining hematopoietic stem cells CD34+CD45dim.

This result is achieved in that in the method of separation of umbilical cord blood leukocyte concentrate containing hematopoietic stem cells, which consists in collecting blood from the umbilical cord into the container, weighing the mass of the collected blood count its volume of a given mass of anticoagulant, the analysis of umbilical cord blood for bacteriological seeding, the implementation of separation by centrifugation, separating umbilical cord blood erythrocyte mass and leukocyte concentrate gemopoeticheskoi stem cells through sedimentologica tools solution hydroxyethylamine, after weighing container, made in the form of a package, with umbilical cord blood in a laminar box irradiated with ultraviolet radiation, selected from a batch of 1.5 ml umbilical cord blood for examination it on geoanalytical, specifying the type of human leukocyte antigen for matching histocompatibility, in the absence or presence of infection, again weigh the mass of the cord blood in the package and determine its volume according to the formula V1=((MPC-Mak)/Pcp)+Vakwhere MPC- weight cord blood, g, with the package in which it is located, MAK- weight anticoag is Lanta, g, with the package in which it is housed, Pcp- the average density of blood, g/ml, Vak- the volume of anticoagulant, ml, and anticoagulant taken in the form of citrate-phosphate-dextrose-adenine with a mass equal to 54.4 g, together with package in which it is housed, the average density of blood is taken equal to 1.05 g/ml, and the volume of anticoagulant - 35 ml after stirring cord blood with anticoagulant, included in the package with umbilical cord blood, in the last injected sedimentologie means in the form of a 6% solution of hydroxyethylamine in the amount calculated by the formula V2=0,2V1placed the package with a mixture of umbilical cord blood, anticoagulant and hydroxyethylcellulose on a platform shaker, carry out orbital mechanical mixing of the contents of the package at a speed of 30 rpm, tilting the platform of the shaker in different planes to the horizon, and spend the incubation mixture with umbilical cord blood for 10 minutes at room temperature, separating the red blood cell mass by sedimentaries in the batch mixture with umbilical cord blood, then placed in the centrifuge chamber to the separator, is connected thereto on a vertical axis of symmetry of the main pipeline, connecting the latter through the optical sensor to the distribution node, from which sum up the pipelines to the bags for collection of blood components and the package with the mixture, fill ka is ERU mixture of umbilical cord blood, anticoagulant and hydroxyethylamine from the package and perform the separation is placed in the centrifuge chamber mixes with umbilical cord blood anticoagulant and hydroxyethylcellulose by centrifugation speeds ranging from 1700 to 8000 rpm in a period of time from 10 to 40 minutes, separating the contents of the package with umbilical cord blood for red blood cell mass, plasma and leukocyte concentrate and distribute them by means of an optical sensor through the pipelines to their bags, after the separation are selected 0.5 ml fractions leukocyte concentrate to determine the viability and number of hematopoietic stem cells CD34+CD45dimon a flow cytometer, from bags of plasma and erythrocyte mass selected 2.5 ml, mix these doses in vitro and test this mixture on bacterial culture for detection of antibodies to T.pallidum in plasma taken 0.1 ml of plasma, also taken out of the bag with plasma 1 ml of plasma in a test tube labeled "plasma child" for research on bloodborne infections, and out of the bag with erythrocyte mass select 1 ml to determine the blood group and rhesus factor.

The invention consists in the combination of essential features, sufficient to achieve provided by the invention a technical result.

The essential features of the proposed method, the ppsr is giving with known characteristics, are: - the collection of blood from the umbilical cord into the container; (B) weighing the mass of the collected blood count its volume of a given mass of anticoagulant; - conduct analysis on bacteriological sowing; G - implementation separation by centrifugation, separating umbilical cord blood erythrocyte mass and leukocyte concentrate from hematopoietic stem cells through sedimentologica tools solution hydroxyethylamine.

Salient features of the process of separation from the umbilical cord blood leukocyte concentrate containing hematopoietic stem cells are: D - after weighing container, made in the form of a package, with umbilical cord blood in a laminar box irradiated with ultraviolet radiation, selected from a batch of 1.5 ml of umbilical cord blood for examination it on geoanalytical, specifying the type of human leukocyte antigen for matching histocompatibility, in the absence or presence of infection; E - again weigh the mass of the cord blood in the package and determine its volume according to the formula V1=((Mnk-Mak)/Pcp)+Vakwhere MPC- weight cord blood, g, with the package in which it is located, MAKis the mass of an anticoagulant, g, with the package in which it is housed, Pcp- the average value of the density to the JVI, g/ml, VAK- the volume of anticoagulant, ml, and anticoagulant taken in the form of citrate-phosphate-dextrose-adenine with a mass equal to 54.4 g together with the package in which it is housed, the average density of blood is equal to 1.05 g/ml, and the volume of anticoagulant - 35 ml; G - after stirring cord blood with anticoagulant, included in the package with umbilical cord blood, in the last injected sedimentologie means in the form of a 6% solution of hydroxyethylamine in the amount calculated by the formula V2=0,2V1; And put the package with a mixture of umbilical cord blood, anticoagulant and hydroxyethylcellulose on a platform shaker, carry out orbital mechanical mixing of the contents of the package at a speed of 30 rpm, tilting the platform of the shaker in different planes to the horizon; To conduct the incubation mixture with umbilical cord blood for 10 minutes at room temperature, separating the red blood cell mass by sedimentaries in the batch mixture with umbilical cord blood; L - place centrifuge chamber to the separator, is connected thereto on a vertical axis of symmetry of the main pipeline, connecting the latter through the optical sensor to the distribution node, from which sum up the lines to the bags collection of blood components and package with the mixture, fill the chamber with a mixture of umbilical cord blood, anticoagulant and hydroxyethylamine package; M - syshestvyut separation placed in the centrifuge chamber mixes with umbilical cord blood at speeds ranging from 1700 to 8000 rpm in a period of time from 10 to 40 minutes, separating umbilical cord blood red cell mass, plasma and leukocyte concentrate and distribute them by means of an optical sensor through the pipelines to their bags; after the separation are selected 0.5 ml fractions leukocyte concentrate to determine the viability and number of hematopoietic stem cells CD34+CD45dimon a flow cytometer; from bags of plasma and erythrocyte mass selected 2.5 ml, mix these doses in vitro and test this mixture on biological seeding; P - for the detection of antibodies T.pallidum in plasma taken 1 ml plasma, P is out of the bag with the selected plasma 1 ml of plasma in a test tube labeled "plasma child" for research on hemotransmissive infection; With - out bag with erythrocyte mass select 1 ml to determine the blood group and rhesus factor.

The method of separation of umbilical cord blood leukocyte concentrate containing hematopoietic stem cells, as follows. From the umbilical cord of the child, separated in physiological delivery of its vein puncture with a syringe and collect cord blood in a special sterile container having input and output ports, made of plastic material, designed for the storage of umbilical cord blood. Record all data of mother and child in the Protocol identification is brazza umbilical cord blood. All further steps will produce a laminar box ("thermo Scientific" Hera Safe KS Heraeus, USA), representing a closed Cabinet with transparent walls and holes in one wall so that you could put your hands in the gloves, using ultraviolet radiation lamp 15 W firm "OSRAM" for decontamination of internal volume. Cloth moistened with 70% acylovir alcohol, wipe the bag with cord blood and the package with the anticoagulant. The anticoagulant is selected in the form of citrate-phosphate-dextrose-adenine with a mass equal to 54.4 g, together with package in which it is placed. Weigh the package with umbilical cord blood in precision electronic scales "ACCULAR" VIC 120d3 RS-232 - VICON USA. After stirring umbilical cord blood kit (manual action) is withdrawn from the package through its port 1.5 ml of umbilical cord blood for her research on geoanalytical and HLA typing, specifying the type of human leukocyte antigen for matching histocompatibility, in the absence or presence of infection. Again weigh the package with a mass of umbilical cord blood and calculate the volume V1the collected cord blood according to the formula V1=((MPC-Mak)/Pcp)+Vakwhere MPC- weight cord blood, g, with the package in which it is located, MAKis the mass of an anticoagulant, g, with the package in which it is placed, equal to 54.4 g, R - the average density of blood, g/ml, and equal to 1.05 g/ml, Vak- the volume of anticoagulant, ml, equal to 35 ml After determining the volume V26% of hydroxyethylamino - sedimentaries the means by which settles erythrocyte mass, by the formula V2=0,2V1enter it in the package with umbilical cord blood. Gently shaking hands, mix the packet with the umbilical cord blood and solution hydroxyethylamine and put this package on a platform shaker (mixer) (Multi Bio 3D (Biosan, Latvia, Switzerland)). Using the latest carry out mechanical orbital mixing of the contents of the umbilical cord blood, anticoagulant and hydroxyethylcellulose at a speed of 30 rpm, tilting the platform of the shaker in different planes to the horizon, the angle 7°, uniformly rocking platform shaker around the coordinate axes - (3 degrees of freedom). Incubated package contents for 10 minutes at room temperature (+18-20°C). Using hydroxyethylamine in the package settled erythrocytes. Centrifuge the camera 1 is installed in the separator 2 (made even more, Switzerland), connected thereto on a vertical axis of symmetry of the pipeline 3, by connecting the latter through the optical sensor 4 to the distribution node 5, which sum up the pipes 6 to the bags 7, 8 for collecting blood components and the package 9 mix 10. Fill the centrifuge chamber 1 from the package 9 a mixture of 10 umbilical cord blood, anticoagulant and hydroxyethylamine. Close the centrifuge chamber 1 and are posted separation in the centrifuge chamber 1 mixture with umbilical cord blood, anticoagulant and hydroxyethylcellulose, centrifugation, i.e. the rotation of the camera around the vertical axis with rotation speed 1700-8000 rpm during the time from 10 to 40 minutes, separating umbilical cord blood plasma, erythrocyte mass and leukocyte concentrate. The speed and time of rotation can be varied depending on the package weight from umbilical cord blood. Through the main pipe 3, through the optical sensor 4 and the distribution unit 5 plasma, erythrocyte mass and leukocyte concentrate distribute its bags 7, 8, 9. Erythrocyte mass fill empty from a mixture of the package bag 9. After separation disconnect each bag from its pipelines and sealed the holes of their pipelines. In laminar box select syringe 0.5 ml leukocyte concentrate in the bullet microprobing to calculate the viability and number of hematopoietic stem cells CD34+CD45dimon a flow cytometer. Out of the bag with a plasma selected for research 5 ml of plasma in a sterile labeled test tube. Of this amount, take 2.5 ml of plasma and from another bag - 2.5 ml of red blood cell mass) is t these doses and test this mixture on bacterial culture. For the test to detect antibodies T.pallidum in plasma taken in a test tube Eppendorf 0.1 ml of plasma. In the bullet test tube plasma of child selected 1 ml plasma to test for bloodborne infection. Out of the bag with erythrocyte mass select 1 ml to determine the blood group and rhesus factor.

Therefore, before storing hematopoietic stem cells in the leukocyte concentrate, derived from umbilical cord blood, they are subjected to bacteriological and virological monitoring for the presence of fungal infective material. The resulting processing of umbilical cord blood material (plasma, erythrocyte mass and leukocyte concentrate) is examined for the determination of blood group and rhesus factor are tested for sterility, viability, identifying a number of infections: HIV, hepatitis b and C, syphilis, determination of the number of hematopoietic stem cells CD34+CD45dim.

The use of technical solutions "isolation from umbilical cord blood leukocyte concentrate containing hematopoietic stem cells in comparison with the prototype, allows to improve the quality and reliability of infection and biogarantie in identified and collected in leukocyte concentrate hematopoietic stem cells, due to the fact that the umbilical cord blood test definition wide-angle on the type of human leukocyte antigen to determine histocompatibility, in the absence or presence of infection plasma isolated from umbilical cord blood, determination of test antibodies T.pallidum, determine on bloodborne infections, testing for virological control. The selection of hematopoietic stem cells hold on effective high quality certified equipment manufacturers in various countries: Switzerland, USA, Denmark, Germany, etc. the process of selection of hematopoietic stem cells carry out commercial organization "LLC Pokrovsky stem cell Bank, which has a license No. 78-01-000356 from 10.10.2008, and # FS-99-01-005911 dated 20.01.2009, on the basis of state Pokrovskaya hospital in Saint-Petersburg. All the technology used in the organization, comply with international requirements and standards for banks stem cells derived from umbilical cord blood. Currently, about 80 different diseases, including cancer, leukemia, lymphoma and specific disorders of the immune system, give a positive response to treatment with stem cells.

The method of separation of umbilical cord blood leukocyte concentrate containing hematopoietic stem cells, which consists in collecting blood from the umbilical cord into the container, weighing the mass of the collected blood count its volume of a given mass of anticoagulant, carrying the AI analysis of umbilical cord blood for bacteriological seeding, the implementation of the separation by centrifugation, separating umbilical cord blood erythrocyte mass and leukocyte concentrate from hematopoietic stem cells through sedimentologica tools solution hydroxyethylamine, characterized in that after weighing container made package, with umbilical cord blood in a laminar box irradiated with ultraviolet radiation, selected from a batch of 1.5 ml of umbilical cord blood for examination it on geoanalytical, specifying the type of human leukocyte antigen for histocompatibility matching in the absence or presence of infection, again weigh the mass of the cord blood in the package and determine its volume according to the formula V1=((MPC-MAK)/Pcp)+VAKwhere, MPC- weight cord blood, g, with the package in which it is located, MAKis the mass of an anticoagulant, g, with the package in which it is housed, Pcf- the average density of blood, g/ml, VAK- the volume of anticoagulant, ml, and anticoagulant taken in the form of citrate-phosphate-dextrose-adenine with a mass equal to 54.4 g together with the package in which it is housed, the average density of blood is equal to 1.05 g/ml, and the volume of anticoagulant - 35 ml after stirring cord blood with anticoagulant, included in the package with umbilical cord blood, in the last injected sedimentologie means in the form of a 6%aqueous solution of hydroxyethylamine in volume, calculated by the formula V2=0,2V1placed the package with a mixture of umbilical cord blood, anticoagulant and hydroxyethylcellulose on a platform shaker, carry out orbital mechanical mixing of the contents of the package at a speed of 30 rpm, tilting the platform of the shaker in different planes to the horizon, and spend the incubation mixture with umbilical cord blood for 10 min at room temperature, separating the red blood cell mass by sedimentaries in the batch mixture with umbilical cord blood, then placed in the centrifuge chamber to the separator, is connected thereto on a vertical axis of symmetry of the main pipeline, connecting the latter through the optical sensor to the distribution node, from which sum up the pipelines to the bags for collection of blood components and the package with the mixture, fill the chamber with a mixture of umbilical cord blood, anticoagulant and hydroxyethylamine from the package and perform the separation is placed in the centrifuge chamber mixes with umbilical cord blood, anticoagulant and hydroxyethylcellulose by centrifugation speeds ranging from 1700 to 8000 rpm in a period of time from 10 to 40 min, separating the umbilical cord blood red cell mass, plasma and leukocyte concentrate and distribute them by means of an optical sensor through the pipelines to their bags, after the separation are selected 0.5 ml fractions leukocyte the CSOs concentrate to determine the viability and number of hematopoietic stem cells CD34+CD45 dimon a flow cytometer of the bags of plasma and erythrocyte mass selected 2.5 ml, mix these doses in vitro and test this mixture on bacterial culture for detection of antibodies to T.pallidum in plasma taken 0.1 ml of plasma, also taken out of the bag with plasma 1 ml of plasma in a test tube labeled "plasma child" for research on bloodborne infections, and out of the bag with erythrocyte mass select 1 ml to determine the blood group and rhesus factor.



 

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31 cl, 4 dwg, 16 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology, and can be used in treatment of mammary gland cancer (MGC). Method is realised in the following way. After confirmation of malignant character of tumour tissue in patient sampling of 200 ml of blood from peripheral vein into reservoir with hemopreservative is carried out, after that it is centrifuged with separation of plasma, which is combined with cyclophosphane in amount 400 mg/m2, doxorubincin in dose 40 mg/m2 is introduced into erythrocyte mass; both reservoirs are incubated in thermostat for 30 minutes at temperature 37°C. After that, sucking of blood from the place of tumour tissue sampling is carried out with further introduction into it of 200 mg of cyclophosphane diluted in 5 ml of physiological solution. Tissue of mammary gland around tumour is infiltrated with incubated plasma with cyclophosphane, and erythrocyte mass with doxorubicin is introduced intravenously by drop infusion.

EFFECT: application of the invention makes it possible to increase efficiency of MGC treatment due to creation of maximal concentration of chemical preparations in leision focus and prevention of tumour process dissemination.

1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medications and deals with application of miliacin as medication which increases antitumour effect of methotrexate.

EFFECT: extension of arsenal of medications, which increase antitumour effect of methotrexate.

3 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly - to ophthalmology. The method involves laser exposure and retinalamine administration For this purpose, retinalamine 5 mg is dissolved in 2% lidocaine 1.5 ml. Parabulbarly, the prepared solution is introduced by 75 ml into each eye. 5-10 minutes later, retina focused continuous noncontact exposure to helium-neon laser follows. The light guide diametre is 3 mm, the distance of a distal end of the light guide to a front surface of cornea is 2-4 cm. The power is 2.2 - 2.6 Wt. Said combined exposure is daily for ten days. The duration of first three sessions of laser exposure is 3 minutes, and the following seven sessions of laser exposure last for 5 minutes.

EFFECT: method improves visual functions due to enhancing visual acuity and extending fields of vision, provides continuous stable maintenance of the effect; it is atraumatic.

3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, specifically to ophthalmology, and can be used in experimental ophthalmology for preparing proliferating crystalline lens cell culture with preferential epithelial cell content for future screening analysis of various methods for prevention of posterior capsule opacity in vitro. Substance of the invention implies mechanical release of a crystalline lens capsule with adjacent cortical mass of a nucleus, refinement to 1 mm3 and processing with mixed 0.05% collagenase first type and 0.25% trypsin; then cell culture is washed by centrifugation with dissolving precipitated cells in a serum-medium DMEM/Ham's F-12; prepared culture is sowed in culture cups in concentration 5×105 cells per cm3 and grown in a CO2 incubator.

EFFECT: advantage of the invention consists in development of a simple method for preparing crystalline lens cell culture preferentially consisting of epithelial cells.

1 cl

FIELD: chemistry.

SUBSTANCE: mixture of chondroitin-6-sulphate and dermatan sulphate extracted from umbilical cords is oxidised with 2,2,6,6-tetramethylpiperidine-1-oxyl-NaClO-NaBr at pH 10.2 and temperature 0-5°C for 1 hour with subsequent precipitation of oxidation products with three ethanol objects and their separation on a chromatographic column with DEAE-cellulose using eluent in form of aqueous solutions of NaCl with increasing concentration 0.2→0.6 M in increment of 0.05.

EFFECT: obtaining pure chondroitin-6-sulphate; the rest of the products are hybrid polysaccharides consisting of chondroitin-6-sulphate and dermatan sulphate oxidised on the primary groups.

1 ex

FIELD: veterinary.

SUBSTANCE: method consists in introducing of mesenchymal stem cells of umbilical cord or placenta, taken after normal childbirth. Endolymphatic introduction of cells is performed by injections of 1-2 ml of physiological solution, which contains 0.075-0.20×106 cells/kg of weight along spine or leg in single-step1-2 times per year.

EFFECT: method allows to improve animal's condition, increasing of its activity, normalises metabolism.

1 ex

FIELD: veterinary.

SUBSTANCE: method consists in introduction of mesenchymal solution of stem cells from umbilical cord or placenta with concentration 0.05-0.250×106 cells/kg of animal weight. At the same time phased hypodermic introduction of one part of stem cells solution in area of metatarsus, tarsus, and saltatory joint of hind legs and two parts of solution in area of sacrum and along lumbar spine is carried out.

EFFECT: method allows increasing of treatment of dog's paralysis and epilepsy, as result of post infection genesis.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to ophthalmology, and aims at treatment of early cataract. A neutral bath of low-molecular peptide fraction that is prepared of acidic protein fraction of muscle extract of mammal lenses fractionated in 100% ammonium sulphate (Vilenzin) is instilled in conjunctival cavity. Instillations are carried out at 9, 12 and 17 o'clock, 2 drops in both eyes twice every 5 minutes within course 2-6 months every 10 days after 30 days.

EFFECT: method provides partial resorption and prevents progression of cataract.

2 ex

FIELD: medicine.

SUBSTANCE: invention concerns medicine, particularly otolaryngology, and can be applied in treatment of acute sensorineural hearing loss. Method involves intramuscular injection of cortexine in amount of 10 mg dissolved in 2 ml of physiological solution, along with vascular and hormonal therapy. Cortexin injection is performed daily for 8-10 days.

EFFECT: enhanced efficiency of acute stage treatment due to complex therapy involving medication combining neuropeptide, nootropic, antioxidation and sedative effects stimulating regeneration process in central and peripheral parts of acoustic analyser.

3 ex

FIELD: medicine.

SUBSTANCE: according to the first version, method includes extraction of allogenic mesenchymal stem cells culture from umbilical cord of newborn after normal delivery for their parenteral introduction in patient according to specified technology. According to the second version, method includes extraction of allogenic mesenchymal stem cells culture from placenta of newborn after normal delivery for their parenteral introduction in patient according to the same technology. The first and second preparations contain the following amount of allogenic mesenchymal stem cells in sterile physiological solution - in the range from 1 to 5 million cells in 5 ml of solution.

EFFECT: increase of therapeutical effect.

6 cl, 2 ex

FIELD: medicine; dermatology.

SUBSTANCE: antimycotic griseofulvin, sunflower seed oil and ascorbic acid are introduced per os. Powder of kerakol from crushed native cornea of neat cattle eyes is applied on infiltrative suppurative focus in amount of 25 mg 2 times per day for 7-8 days.

EFFECT: formation of tender cosmetic scar in focus of infiltrative suppurative trichophytosis, method simplification and reduction of treatment time.

2 ex

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