The method of obtaining immunostimulant

 

(57) Abstract:

The method is designed to obtain immunostimulant as a food additive. The method consists in the following. Isolated bone marrow cells from the bone marrow of small cattle, washed and incubated them in oncotic solution with a concentration of 0.85 - 0.90%, and then centrifuged supernatant. The method allows to extend the resource base, to accelerate and reduce the cost of obtaining immunostimulant as a food additive. table 1.

The invention relates to the field of food industry, in particular to methods for food additives.

A method of obtaining extracts for medical purposes, having an immunostimulating action, from raw materials of animal origin by isolating bone marrow, heat free from pyrogens water and keeping at 80oC for 5 min, centrifuge and use the supernatant for subsequent ultrafiltration, electrodialysis (see and.with. N 1442064, MKI A 61 K 37/24, 1988, BI N 44).

However, the known method is difficult to implement and economically unfeasible to obtain immunostimulant in the form of nutritional supplements.

Enableuser by separating the cells from the bone marrow of animals, followed by incubation in growth medium, the preconcentration and elution supernatant (see and. C. N 1005790, MKI A 61 K 39/00, 1983, BI N 11).

However, the known method is time-consuming, because it requires strict sterile conditions, expensive due to the use of nutrient medium 199, which includes the following components (g/l):NaCl-8, KCl-0.4, MgCl26H2O-0.1, MgSO47H2O-0.1, Na2HPO47H2O-0.06, KH2PO47H2O-0.06, CaCl2-0.14, NaHCO3-0.6, fanart-0.02, glucose-0.6, the application of which does not allow to use it for food purposes. The method involves the use of only ordinary bones of pigs.

Thus, the problem to be solved in the present invention, is to obtain immunostimulant in the form of dietary supplements. The technical result obtained by carrying out the method is a simplification of the process and the expansion of raw materials.

To achieve provided by the invention of the technical result in the production method of immunostimulant by separating the bone marrow cells of animals, followed by incubation in growth medium, centrifugation supernatant, according to the invention, the cells isolated from the bone marrow of small horn the entrances take in a given concentration.

As a result of experiments, the authors showed the possibility of using oncotic solution with a concentration of 0.85 - 0.90% as a nutrient medium for the cells in the bone marrow of animals. The authors found that you can use not only the bone marrow cells of the pigs and small ruminants. The cell concentration was chosen so as to obtain the maximum yield of biologically active components and to maintain cell viability.

Analysis of patent and scientific literature showed that of the prior art, the authors have not identified technical solutions, including a collection of characteristics that are similar or equivalent to the claimed.

This allows to make a conclusion on the conformity of the proposal with the criteria of "novelty" and "inventive step".

The proposed method for immunostimulant is as follows.

When the butchering of carcasses at the slaughterhouse retrieved ribs of animals, it can be ribs pigs or small cattle. Washed cells from the rib bones produce syringe with oncotic solution with a concentration of 0.85 - 0.90%. Oncotic solution is a natural environment for Iznik selection, and incubation, without compromising the integrity of the cell. Then produce three times washing of the cells with the same solution: a suspension of cells centrifuged, the supernatant fluid is drained off, and the residue diluted oncotic solution. In the last set concentration by counting cells per 1 ml oncotic solution in the chamber Goryaeva. In order to achieve a given concentration of cells after counting the produce or concentration, or dilution of the suspension of cells oncotic solution. What is the cell concentration in the inventive method of obtaining immunostimulant the authors do not specify, since it is the "know-how" in the present invention. And the authors do not wish this moment was uncovered. At this concentration, the cells are placed in culture bottles of 100 - 150 ml of sediment in the bottle, the volume of one litre. Incubation is carried out at a temperature of 37oC for 14 to 18 hours after incubation, a suspension of cells centrifuged, the supernatant determine the amount of protein by the method of Lowry, and then either used in the experiment, or stored at a temperature of -12oC.

Evaluation of immunological activity derived immunostimulant conducted in vi is private run.

Example 1. When the butchering of carcasses at the slaughterhouse remove ribs pigs. Cells wash away from the rib bones of pigs by fioratura, washed and incubated in the specified concentration for 16 hours at a temperature of 37oC in the culture vials of 100 - 150 ml of suspension in a vial, the volume of which 1 liter of the Supernatant liquid is separated from the cells by centrifugation at 2000 minutes the supernatant determine the amount of protein by Lowry. The content of 2 mg/ml

Evaluation of immunological activity performed in the system in vivo for the local introduction of faction mice at 4 days secondary immune response to sheep erythrocytes in a dose of 100 µg contained in 0.1 ml of fidirectory, pad hindquarters. In the other group, which served as a control, enter fisierului. The next day in these mice, remove the popliteal lymph nodes and define them in the number of antibody productive cells, comparing their number in the experiment and the control.

The coefficient of stimulation of antibody productions To determine in both models calculated by the ratio of the number of the KLA in the experience to the corresponding number of the KLA in control.

Example 2. When the butchering of carcasses at the slaughterhouse remove ribs small ncentratio for 16 hours at a temperature of 37oC in the culture vials of 100 - 150 ml of suspension in a vial, the volume of which 1 liter of the Supernatant liquid is separated from the cells by centrifugation at 2000 minutes the supernatant determine the amount of protein by Lowry. The content of 2.1 mg/ml together with the selection of cells with medium 199, washing and incubation are in the same environment. Conditions for obtaining analogous to example 1. The protein content of 2.5 mg/ml

Evaluation of immunological activity performed in the system in vivo for the local introduction of faction mice at 4 days secondary immune response to sheep erythrocytes. In the case of obtaining compositions with medium 199 in the group served as control, injected with medium 199.

The coefficient of stimulation of antibody productions To determine in both models is calculated as the ratio of the number of KLA in the experience to the corresponding number of the KLA in control.

Example 3. When the butchering of carcasses at the slaughterhouse remove ribs pigs. Cells wash away from the rib bones of pigs by fioratura, washed and incubated in the specified concentration for 16 hours at a temperature of 37oC in the culture bottles. The supernatant liquid is separated from the cells by centrifugation at 2000 minutes the supernatant operadelaware to use products, subjected to heat treatment, and to maintain its biological activity, the supernatant is heated at 100oC for 10 minutes

Evaluation of the immunological activity of the composition subjected to heating, is carried out in the system in vivo is similar to the first two examples. The coefficient of stimulation of antibody productions To determine in both models calculated by the ratio of the number of the KLA in the experience to the corresponding number of the KLA in control.

Example 4. When the butchering of carcasses at the slaughterhouse remove ribs small cattle. Cell wash, washed and incubated with medium 199. Incubation at a given concentration carried out for 16 h at a temperature of 37oC in the culture bottles. The supernatant liquid is separated from the cells by centrifugation at 2000 minutes the supernatant determine the amount of protein by Lowry. The content of 2.8 mg/ml

When choosing the ways of preserving food compositions considered cooling at 0 to 4oC, freezing at -18oC and storage at -12oC for 1 month. Cooling was not effective due to rapid contamination, and the fraction subjected to freezing and storage at Racchi mice at 4 days secondary immune response to sheep erythrocytes.

The technical nature of the claimed invention was compared with the prototype (see and.with. N 1005790, class A 61 K 39/00, 1983, BI N 11).

The coefficient of stimulation of antibody productions To determine in both models calculated by the ratio of the number of the KLA in the experience to the corresponding number of the KLA in control.

Data examples are given in the table.

Thus, the proposed method allows to obtain bone marrow cells not only from the bones of pigs, but the bones of small cattle, which can significantly extend the raw materials for the production of dietary supplements. Immune obtained by this method, retains its biological properties both at high and at low temperatures. Compared with the known method of producing stimulator antitelomerase the proposed method for obtaining reduced by 2 to 4 h; simplified due to the refusal of the gel filtration, lyophilization; replacement of the nutrient medium 199 oncotic solution significantly reduces the cost of the method, the composition obtained in this way can be stored at low temperatures.

The immunostimulant in the form of dietary supplements will allow you to get the products for spamai pesticides and food additives are well known formulation of meat products with the introduction of the proposed immunostimulant.

The method of obtaining immunostimulant by separating the cells from the bone marrow of animals, followed by incubation in growth medium, centrifugation of the supernatant liquid, characterized in that the cells isolated from the bone marrow of small cattle, as well as the nutrient medium used oncotic solution with a concentration of 0.85-0.90%, and the cells take in a given concentration.

 

Same patents:

The invention relates to medicine, namely General medicine and dentistry

The invention relates to medicine, namely to methods for preparations from bone marrow, in particular negative regulators of hemopoiesis
The invention relates to medicine, in particular for surgical treatment of musculoskeletal system
The invention relates to medicine, namely the creation of an organ-specific homeopathic drugs for stimulation of myeloid and lymphoid blood sprouts
Solution: bone // 2055535
The invention relates to medicine, namely to traumatology and orthopedics, dental surgery, and can be used in the treatment of patients with lesions of bone

The invention relates to the production of the drugs used in medical practice, and relates to a method of obtaining domestic product, similar to the mechanism of biological action of the drug "Cerebrolysin" firm Ebewe, Austria
The invention relates to the production of the drugs used in medical practice, and relates to a method of obtaining domestic product, similar to the mechanism of biological action of the drug "Cerebrolysin" company Ebewe, Austria

The invention relates to medicine, in particular, to a method for immunomodulator of mielopid

FIELD: medicine, surgery, transplantology.

SUBSTANCE: embryonic spleen should be sampled, washed in nutritive medium № 199 to be placed into fresh medium № 199 to obtain homogenate in teflon homogenizer followed by centrifuging; then one should isolate the upper, medium and inferior layers, suck off medium layer and the upper part of inferior layer; the cell mixture obtained should be diluted in nutritive medium № 199 to be then introduced by injections into mesentery of small intestine or rectus muscle of abdomen. The present innovation favors the activation of immune system in patients undergone splenectomic operation and in those in case of surgical immunodefficient state due to high functional and regenerating activity of transferred embryonic splenic cells.

EFFECT: higher efficiency of prophylaxis.

6 dwg, 2 tbl

FIELD: medicine.

SUBSTANCE: method involves using mononuclear autologic marrow fraction containing 6-9x104 mesenchyma cells per 1 ml or autologic mesenchyma trunk cells. The cells are separated from brain bioptate in the amount of 106 cells/kg of patient body mass. The preparations are intracoronarily introduced in fractions at a rate of 3-5 ml/min into the right coronary artery. The introduction is also carried out in intra-arterial mode in jets or in drops.

EFFECT: higher survival rate and life quality of cardiologic patients.

1 dwg

FIELD: medicine.

SUBSTANCE: method involves introducing autologic mesenchyma trunk cells as a single intravenous drop dose or in the amount of 1 mln cells per 1 kg of patient body weight.

EFFECT: provided stable clinical remission.

FIELD: medicine, surgery.

SUBSTANCE: method involves every day covering a wound with an agent comprising of pig spleen homogenate and oxygenated with perfluorane taken in the following amounts: 100 g of pig spleen homogenate and 10 ml of oxygenated perfluorane. Invention promotes to accelerating sanitation of suppurative wounds due to activation of topical immunity and activation of regenerating processes. Invention can be used in treatment of suppurative wounds.

EFFECT: improved treatment method of wounds.

2 ex

FIELD: medicine, oncourology.

SUBSTANCE: the present innovation deals with treating locally spread tumor diseases of urinary bladder due to applying either chemo- and/or radiation therapy. Moreover, one should intravenously once inject by drops autologous mesenchymal stem cells (AMSC) at the quantity of 1 mln cells/kg patient's body weight. Moreover, in case of chemotherapy AMSC should be injected during the period before the 20-th d after the last injection of chemopreparation, in case of radiation therapy - 12-15 d against the day of irradiation. In case of chemoradiation therapy AMSC should be injected after chemotherapy before the course of radiation therapy. This method enables to carry out chemoradiation therapy in patients with severe accompanying diseases that prevent such a therapy, and prevent the development of general toxic complications of chemoradiation therapy and medicinal and radiation-caused cystitis.

EFFECT: higher efficiency of therapy.

3 ex, 2 tbl

FIELD: medicine; medical engineering.

SUBSTANCE: biotransplant has genetically unmodified mesenchyma stem cell culture as active component obtained from fetal donor autologous material. The tissue is subjected to disaggregation and the produced cell suspension is resuspended and cultivated on growth medium containing transferrin, insulin, fibroblast growth factor and heparin to accumulate mature stroma in cell culture. Method involves intravenously dropping mesenchyma stem cell culture in the amount of 50 to 500 mln in 50-100 ml of physiologic saline.

EFFECT: accelerated recovery of bone tissue; positive biochemical factors dynamics; improved patient locomotor activity.

6 cl

FIELD: medicine; medical engineering.

SUBSTANCE: biotransplant has genetically unmodified mesenchyma stem cell culture as active component obtained from fetal donor autologous material and genetically unmodified fetal myoblast culture. Method involves intravenously dropping mesenchyma stem cells in the amount of 50 to 500 mln in 50-100 ml of physiologic saline. The fetal myoblast culture is intramuscularly introduced at a dose of 100 mln of cells per 10 kg.

EFFECT: enhanced effectiveness in repairing incretory function of pancreas and reducing resistance to insulin; reduced risk of nephropathy and other complications.

9 cl

FIELD: medicine, cardiology.

SUBSTANCE: one should introduce the suspension of autologous mononuclear medullary cells without preliminary cultivation in to the mouth of coronary artery nourishing infarction area. Cell suspension should be introduced at the quantity of 100-150 mln. cells immediately after stenting coronary artery due to technique of passive passage. The procedure should be performed on the 14th - 21st d against the onset of the disease mentioned. The method provides efficient counterbalance of cardiomyocytes due to applying valuable stem cells at excluding complications induced by arterial occlusion.

EFFECT: higher efficiency of therapy.

1 ex, 1 tbl

FIELD: medicine, in particular hematology.

SUBSTANCE: CBA/CaLac intact mouse spleen is homogenized in medium containing 95 % of RPMI-1640 and 5 % of ethyl silicate; filtered through nylon grid; washed by centrifugation with RPMI-1640. at 1500 rpm for 10-15 min; final concentration of living splenocites is adjusted to 2x106 colony/ml using media containing 10 % of ethyl silicate, 1 % of L-glutamine; 0.2 % of gentamycine, 0.05 % of mercaptoethanol, 89 % of RPMI-1640; obtained product is cultivated at 37°C for 6 days, centrifuged and supernatant containing target product is separated followed by storage at -20°C. Before filtering 1 ml of distilled water is added in sterile spleen homogenate and after 25 sec 10 ml of RPMI-1640 medium is added. Target product is stored for 1 month or less.

EFFECT: method of improved accuracy and validity.

1 tbl

FIELD: medicine, in particular uses of viruses for cell elimination.

SUBSTANCE: invention relates to uses of viruses capable of reducing undesired cells in ex vivo mixtures of normal marrow or peripheral blood cells and tumor cells, such as leucosis or lymphoma cells due to interaction of abovementioned mixture with vesicular stomatitis virus. Invention also relates to method for malignant tumor treatment in mammalian. Claimed method includes sampling of mammalian marrow or peripheral blood cells; interaction of said cells ex vivo with vesicular stomatitis virus; mieloablative therapy; and transplantation of purified hematopoietic cells into mammalian. In another embodiment invention relates to method for malignant tumor treatment in mammalian having transplant of marrow or peripheral blood stem cells. Said method includes interaction ex vivo of collected transplant cell with vesicular stomatitis virus and administration of these purified cells in mammalian. Method of present invention makes it possible to reduce risk of malignant tumor backset in mammalian with transplanted hematopoietic cells.

EFFECT: new agent for elimination of marrow or peripheral blood tumor cells.

12 cl, 4 ex, 3 tbl

Up!