Cell line of human melanoma mel h, applied for obtaining antitumour vaccine
SUBSTANCE: cell line of humans melanoma mel H has stable cultural and morphological characteristics, is stored in Specialised collection of cell cultures of vertebrates of Russian collection of cell cultures of Institute of Cytology RAS by number "РККК (П) 715 Д".
EFFECT: invention allows to extend arsenal of human melanoma lines, which are used for creation of antitumour vaccine, applied for treatment of melanoma and other malignant neoplasms.
1 tbl, 2 ex
The invention relates to the field of medical biotechnology, in particular the production of cell lines used to generate antitumor vaccines.
Vaccine therapy is one of immunological approaches to cancer treatment. The principle of this method is based on the induction of antitumor immunity after injection into the body of the tumor antigen.
The Central event in the process of T-cell immune response against tumor cells is to stimulate recognition of the T-receptor antigenic determinants, selectively expressed on tumor cells. Tumor antigens, as a rule, subjected to processing prior to their presentation in the context of histocompatibility molecules on the cell surface. Different categories of tumor-associated antigens can be divided into three main groups: cancer/testicular antigens (MAGE, BAGE, FRAME, NY-ESO-1, HOM-MEL-40), differencirovanie antigens of melanocytes (tyrosinase, Melan-A/MART-1, gp100, TRP-1, TRP-2), and mutated antigens (MUC-1, CDK4, β-catenin gp100-in4, p15, N-acetylglucosaminyltransferase V). With immunologically cancer/testicular antigens may be good targets for immunotherapy of tumors, because in normal tissues, this group antigens (MAGE and FRAME) is not expressed, except in the tissue of the testicles, which half is available for cells of the immune system due to the absence of their direct contact with immunocompetent cells  and lack of expression of HLA antigens class I . Unlike cancer/testicular antigen immunogenicity differencirovannyh antigens of melanocytes low due to immunological tolerance to these "its" antigens. However, this antigen Melan-A/MART-1, contains several epitopes for recognition CTLs (cytotoxic lymphocytes) and is able to induce the generation of melanoma-specific CTLs.
Thus, expression of various tumor markers plays a key role in the induction of antitumor immunity. A variety of relevant antigens allows for more "complementary" to select cell lines to generate antitumor vaccines.
Vaccines prepared on the basis of tumor cells are whole cell vaccines and are living allogeneic or autologous tumor cells.
Autologous/syngeneic solid tumor cells comprise almost all of the antigens expressed by the tumor host, which reduces the risk of allergic reactions to alien neoprogressive antigens, and also reduces the risk of contamination with pathogenic viruses and intracellular parasites. A vaccine consisting of multiple cell lines (polyvalent vaccine), contains a broad spectrum of tumor antigens and is used as allogeneic. Such polyvalent vaccine, as the vaccine Mrtona et al. , consists of three allogeneic melanoma cell lines with high expression of immunogenic surface Glyco - and lipoproteins and gangliosides. Clinical trials of this vaccine have shown that the development of the immune response as the cellular and humoral type, these antigens was correlated with increased survival of patients. Another vaccine, "Melacine"  (Corixa corp., Canada), consisting of lysate allogeneic melanoma cell lines, induces antitumor effect in 5-10% of patients with melanoma.
The present invention is to obtain a new tumor cell lines human melanoma carrying a particular set of antigens that can be used in the creation of antitumor vaccines.
The technical result to be obtained by the use of the invention is expressed in expanding Arsenal of cell lines used to generate antitumor vaccine (whole cell, genetically engineered), which gives the opportunity to increase the effectiveness of treatment and increase life expectancy in the treatment of malignant tumors.
The problem is solved in that the resulting new cell line mel N of the tumor sample disseminated melanoma human skin.
The obtained cell line has a stable cultural and morphological characteristics. X is anicca in cell culture collection of the Institute of Cytology RAS number RCCC(P) D.
Lineage cell line mel H
Cell line derived from a tumor sample of a patient HNS, and/b 00/9865, were treated in 2001 with a diagnosis of disseminated melanoma skin of the left foot. Tumor material obtained by removal of metastatic inguinal lymph node. Previous treatment: surgery, immuno - and chemotherapy. Steadily growing cell line was obtained on 31 passage.
Morphological signs of cell lines mel H
Cell line mel H consists of relatively monomorphic cells of the small and medium size, round, oval and elongated without a clear cytoplasmic loops. The cytoplasm is abundant inhomogeneous, painted in basophilic tone with a pinkish tinge. Cell nuclei Hypo - and normocapnia with granular chromatin structure, sometimes contain nucleoli. Rare 2 nuclear and giant mononuclear and multinucleated cells. There are mitoses.
Karyological characterization of cell lines ml N
The culture is characterized by different diameters of the nuclei at the same density of color and the state of chromatin, characteristic of interphase nuclei. Analyzed 88 metaphases. The number of chromosomes in 69 cells ranges from 54 to 65 chromosomes, reflecting triploid set of (3n). In 19 cells detected hexaploid (6n) set (about 120 km HRO is Osom). This cell with replicated chromosome sets in which there was no separation of the cytoplasm, i.e., the presence of increased polyploidization. Found additional chromosomal material of unknown origin on the long arm of the two chromosomes 19 and on the long arm of chromosome from the group D, the inversion of part of chromosome 16, translocation of chromosomes 13 and 15. In 32 cells observed chromosomal aberrations, presents dicentrine (47), tricentenary (1) and ring (3) chromosomes. Therefore, this culture is characterized by high chromosomal variability. Karyotype - 54~65<3n>,der(13;15)(q10;q10),add(Dq),inv(16)(p11q12),add(19)(q13.4).
Immunogenetic study (determination of HLA phenotypes (class I))
A3, A30(19); B13, B7.
Cultural properties of the cell line ml N
Cell line mel H is cultivated in a nutrient medium RPMI (90%), fetal calf serum 10%, containing antibiotics (penicillin with streptomycin at a concentration of 100 u/ml and 100 µg/ml, respectively). In culture flasks (25 cm2in 5 ml of medium seeded with 1×106cells. The cultivation temperature of 37°C. the Monolayer of cells is formed in 3-4 days. When seeding concentration of 70-100 thousand/ml monolayer is formed at 2-3 days without changing the medium. Cells are removed using standard solutions of 0.25% solution of trypsin and 0.02% solution of Ve is sung in a 1:1 ratio. When planting a concentration of 500 thousand/ml index of cell proliferation after 48 hours of cultivation is 3.6-4.6.
Conditions of cryopreservation
For long-term storage of cells preserved by freezing in liquid nitrogen. Cells resuspended environment for freeze - nutrient medium RPMI (80%), fetal calf serum 20%, 10%DMSO. The mode of freezing: liquid nitrogen, the temperature reduction of 1°C per minute up to minus 25°C, then rapid freezing to minus 70°C. Storage in liquid nitrogen at minus 196°C. Defrost quickly, at 37°C. Cells diluted in 10 ml serum-free medium and precipitated by centrifugation, resuspended in 5 ml of the same medium containing 10% fetal calf serum, and transferred into a culture flask with a volume of 25 see cell Viability is assessed according to the inclusion Trypanosoma blue. Cell viability after thawing is 90%.
With long-term observation of bacteria and fungi in culture is not detected. Test for Mycoplasma negative.
Examples of the use of cell lines ml N
Example 1. Cultivation of cell lines mel H. Tumor tissue obtained during surgical removal of melanoma metastases were divided mechanically into fragments the size of 2-3 mm3in the medium RPMI-1640, then, using Cell dissociation sieve-tissue kit" (Sigma)is Uchali cell suspension. The number of viable cells was determined by a standard method in the camera Goryaeva, using 0.5% solution Trypanosoma blue in PBS. In culture flasks (25 cm in 5 ml of medium were seeded at 1×106cells. The cultivation temperature of 37°C. Cells were cultured in medium RPMI 1640 containing 10% fetal calf serum, 2 mm L-glutamine, 1% HEPES, penicillin (100 u/ml), streptomycin (100 μg/ml) and a complex of amino acids and vitamins (Flow Lab.) in the culture flasks (Costar). After passage 31 received steadily growing cell line.
Example 2. The determination of antigens located on the cell line ml N. The obtained cell line mel H, with a stable cultural and morphological characteristics, using methods immunofluorescence, immunohistochemistry, PCR (polymerase chain reaction) analysis was investigated on expressed genes (differencirovanie, tumor-associated and histocompatibility). Differencirovanie melanoma markers that define this line to melanoma, was investigated using monoclonal antibodies CD63, NPS, MelanA, Tyrosinaza, HMW. The histocompatibility antigen identified by using monoclonal antibodies in immunofluorescence reaction.
This cell line is characterized by the expression of melanoma (differencirovannyh) markers: HMW and CD63, antigene the histocompatibility first class emphasizing the specificity of this cell line. Negative experse differencirovannyh markers NMW, MelanA, Tyrosinaza. Positive expression of cancer-testicular genes MAGE corresponds cancer profile and can be used for this line to create cancer vaccines. The data obtained is displayed in table 1.
|The expression of antigens on the cell line mel N|
|Differencirovanie antigens||Cancer-testicular antigens||The histocompatibility antigen|
|CD63||I promise.||MAGE||I promise.||HLA (class I)||I promise.|
As follows from the table. 1, this cell line is characterized by the expression of melanoma markers: HMW and CD63, emphasizing the specificity of this cell line. Positive expression of cancer-testicular markers collection MAGE corresponds cancer profile and can be used for this line to create cancer vaccines. Antigens presented by the major histocompatibility molecule of the first class (HLA).
Thus, this cell line melanoma human skin mel N has its own individual phenotype of tumor markers, implying the presence of differencirovannyh antigens - HMW, CD63, cancer-testicular genes of the MAGE class, as well as molecules of the major histocompatibility first class and the absence of such differencirovannyh markers, as MelanA, HMB45, Tyrosinaza. This phenotype allows the use of the cell line to create an antitumor vaccine (whole cell, genetically the engineering), used to treat melanoma and other malignant neoplasms.
1. Barker C.F. et al. Immunologically privileged sites. ADV. Immunol. 1977, 25: 1-54.
2. Tomita Y. et al. Immunohistochemical detection of intracellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex class I antigens in seminoma J. Urol. 149: 659-663, 1993.
3. Morton DL et al. Ann N Y Acad Sci 1993; 690: 120.
4. V.K. Sondak, J.A. Sosman Results of clinical trials with an allogenic melanoma tumor cell lysate vaccine: Melacine/ Semin cancer Biol. 2003. Dec. 13(6): 409-15.
Cell line human melanoma mel H used to obtain anti-cancer vaccines, is stored in a Specialized collection of cell cultures vertebrate Russian cell culture collection of the Institute of Cytology RAS number RCCC (P) D.
SUBSTANCE: cell line of human melanoma mel Me has stable cultural and morphological characteristics, is stored in Specialised collection of cell cultures of vertebrates of Russian collection of cell cultures of Institute of Cytology RAS by number "РККК (П) 712 Д".
EFFECT: invention allows to extend arsenal of cell lines of human melanoma, applied for creation of antitumour vaccine, which are applied for treatment of melanoma and other malignant neoplasms.
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SUBSTANCE: cell line of human melanoma mel Rac has stable cultural and morphological characteristics, is stored in Specialised collection of cell cultures of Institute of Cytology RAS by number "РККК (П) 710 Д".
EFFECT: invention allows to extend arsenal of cell lines of human melanoma, used for creation of antitumour vaccines, which are applied for treatment of melanoma and other malignant neoplasms.
1 tbl, 2 ex
SUBSTANCE: invention refers to medicine and biology, more specifically to technology and methods for recovery of leukaemia cell suspension from spleens of high-leukaemia mice, and can be used in experimental oncological haematology for storage of low-frequent subtypes of murine leukaemia. There is offered a method for preparing a leukaemia cell suspension from spleens of inbred mice AKR/JY involving skin preparation with an antiseptic, splenectomy, decapsulation and homogenisation of spleen, filtration of the prepared suspension to be diluted in a sterile cooled artificial extracellular medium to the required concentration of leukaemia cells, prepackaging of the prepared suspension in ampoules for storage and transportation with decapsulation, grinding and homogenisation of fragmented spleen pulp performed in polyglucin solution containing ozone 25.72 mg/l and cooled to 24.0±1.0°C, leukaemia cells are filtered and diluted in polyglucin solution containing ozone 25.72 mg/l and cooled to 15.0±2.0°C.
EFFECT: invention provides maximum safety of specific functional adequacy of the transplanted leukaemia cells.
1 ex, 1 tbl
SUBSTANCE: invention relates to biotechnology and can be used in treatment of diseases, namely oncologic ones. Method is based on sampling from patient immature dendrite cells, their cultivation ex vivo for maturing and formation of allostimulating activity. Cells are additionally collected with antigen and are introduced to the same patient for formation of adaptive immunity. During cultivation ex vivo of immature dendrite cells fragmented allogenic double-stranded genomic DNA with fragments with size 200-6000 bp is introduced into culture medium in amount 5 mcg/ml of medium.
EFFECT: invention allows to extend field of obtained product application with increase of general immune state of patients.
SUBSTANCE: there is produced a new CT-B1/F8 hybrid cell clone producing monoclonal cholera toxin antibody (MCAB) in the environment of cell culture and abdominal cavity of syngeneic animals. The clone is produced by the fusion of mouse myeloma SP-2/0 cells with popliteal lymph node cells of BALB/c mice immunised with a commercial preparation of cholera toxin (SIGMA) in posterior pads. A fusing agent is polyethylene glycol of molecular weight 4000. Hybridoma selection has been performed on Dulbecco modified Eagle's medium with bovine foetal serum and hypoxantine-aminopterin-thymidine added. Hybridoma synthesises MCAB specifically interacting with cholera toxin and not interacting with thermolabile E.coli toxin. The antibody titre reaches 1:20000 in the cultural fluid, 1:2000000 in ascitic.
EFFECT: antibodies can be used for designing immunobiological systems of cholera toxin detection exceeding available analogues in sensitivity.
2 dwg, 4 ex
SUBSTANCE: there is offered a method of precursor recovery from a human body, including all cells having stem cell characteristics or similar, in particular closely active or multiple active precursors, and such cells are recovered either directly, or indirectly from human breast secretion sampled from the specified human body. This secret can be colostrum, ripened milk or secretions in males and females in a lactation interval, during at least one of the following periods: absence of pregnancy, pregnancy period, lactation period, involution period. Further, the present invention refers to preferable applications of such recovered cells.
EFFECT: improved clinical effectiveness.
24 cl, 8 dwg
SUBSTANCE: invention relates to medicine, namely to methods of obtaining deposited lymphokine-activated killer cells (LAKC) for treatment of oncologic diseases. Claimed is method, according to which mononuclear lymphocytes (MNL) are isolated from peripheral blood of malignant effusion by centrifugation on one-step gradient of ficoll with density 1.06-1.08 g/cm3, LAKC are generated and concentrated by successive precipitation of MNL by centrifugation, re-slurring it in RPMI 1640 or DMEM medium with addition of 2-10% AB human serum, interleikin-2 in concentration 0.5-1.0 mlnIU/ml and incubation in CO2 incubator for 48-72 hours, after that suspension of obtained LAKC is deposited in culture medium in amount 2 mln LAKC per 200 ml of medium on sterilised and washed with culture medium porous titanium carriers with 55-60% porosity.
EFFECT: invention allows to increase efficiency of obtained medication for local and local-regional immunotherapy of oncologic patients.
1 ex, 8 dwg, 2 tbl
SUBSTANCE: diploid cell cultures are reduced and collected by propagation in a monolayer in an Eagle's growth medium. The conditioned growth medium is separated from the formed cell layer by sterile bottling. The cell layer is removed, washed by centrifugation. The prepared cell suspension is reduced to the required and bottled in the sterile environment. The conditioned growth medium bottles and cell suspension bottles are consistently single frozen to temperature -20°C and higher and thawed at temperature +25°C and lower, and bottled in the sterile environment. The conditioned growth medium and cell suspension are frozen in open bottles at temperature minus 50°C and lower, kept in the frozen environment for at least 48 hours and then lyophilised in two stages. The first stage involves desorption at temperature minus 50°C to 0°C for at least 20 hours; and the second stage requires sublimation for at least 20 hours at temperature 0°C to 30°C; the finished lyophilised bottles are closed in the sterile environment. The invention allows preparing the lyophilised preparations effective in treatment of deep vast burning wounds, frostbites, degrees III (a, b) and IV subdermal burns, treatment of oral and nasopharyngeal mucosa, are storage-stable at temperature +4°C for at least 12 months.
EFFECT: higher efficacy of the preparations for treatment of diseases.
1 dwg, 3 tbl, 2 cl
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to genetic engineering and can be used to optimise expression of the antigen protein of the human epidermal growth factor-2 (HER2/neu). To obtain the HER2/neu protein, a nucleic acid synthetic molecule is used, which is codon-optimised for high level of expression of the said protein in a human cell.
EFFECT: invention increases production of the recombinant HER2/neu protein during expression in human cells.
8 cl, 10 dwg, 14 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology, specifically to obtaining cell population and can be used in cell transplantology and tissue engineering in order to obtain cell material for restoring nerve tissue damaged due to injury, stroke or neurodegenerative diseases. The method involves isolating a population of stromal cells from fat tissue, immunosorting based on using antibodies against the brain-derived neurotropic factor (BDNF) receptor (TrkB) to obtain an essentially homogeneous sub-population of TrkB-expressing cells for their culturing in a standard medium for mammal cells and differentiation induction through transfer into a medium which contains a neural differentiation inducing substance - BDNF combined with 5-azacytidine or retinoic acid combined with 5-azacytidine.
EFFECT: invention enables to obtain a population of neural differentiation induced stromal cells of fat tissue.
7 cl, 5 dwg, 4 tbl, 6 ex
FIELD: organic chemistry, natural compounds, medicine, oncology.
SUBSTANCE: invention represents new saponin mixtures used for inhibition of initiation and activation of mammalian epithelial cell in pre-malignant or malignant state, for stimulation of apoptosis of mammalian malignant cell, prophylaxis of anomalous proliferation of mammalian epithelial cell, for treatment of inflammatory and regulation of angiogenesis in mammal. These mixtures are isolated form plants of species Acacia victoriae. Also, invention relates to methods for their applying. These compounds can comprise triterpene component, such as acacic or oleanolic acid to which oligosaccharides and monoterpenoid components are joined. Mixtures and compounds elicit properties associated with regulation of apoptosis and cytotoxicity of cells and strong anti-tumor effect with respect to different tumor cells.
EFFECT: valuable medicinal properties of compositions.
43 cl, 53 tbl, 50 dwg, 44 ex
FIELD: biotechnology, molecular biology, medicine, genetic engineering, pharmacy.
SUBSTANCE: the hemopoietic protein comprises the amino acid sequence of the formula: R1-L1-R1, R2-L1-R1, R1-R2 or R2-R1 wherein R1 represents the modified ligand flt-3; R2 represents the modified human IL-3, the modified or unmodified colony-stimulating factor. Modification of R1 is carried out by addition of N-end with C-end directly or through linker (L2) that is able to join N-end with C-end to form new C- and N-ends. The modified human IL-3 is prepared by replacing amino acids at positions 17-123. The human G-CSF is modified by exchange of amino acids. The hemopoietic protein is prepared by culturing cells transformed with vector comprising DNA that encodes the hemopoietic protein. The hemopoietic protein stimulates producing hemopoietic cells and this protein is used as a component of pharmaceutical composition used in treatment of humans suffering with tumor, infectious or autoimmune disease. Invention provides preparing multifunctional hemopoietic proteins eliciting the enhanced activity with respect to stimulation of hemopoietic cells and eliciting the improved physical indices. Invention can be used for preparing chimeric multifunctional hemopoietic proteins.
EFFECT: improved preparing and producing method, valuable medicinal properties of protein.
22 cl, 19 dwg, 18 tbl, 117 ex
FIELD: cellular biology, medicine.
SUBSTANCE: invention relates to isolating and cryopreserving precursor-cells. Methods involve treatment of human liver tissue for preparing the essentially monocellular suspension containing precursor-cells and cells that are not precursor-cells, a single or more lines of cellular differentiation presenting in the human liver. Invention describes methods involving stage for separating cellular population resulting to reducing amount of cells that are not precursor-cells and providing preparing the separated suspension enriched with precursor-cells expressing one or more markers and associated with a single or more lines of the cellular differentiation. Also, invention describes a method for selection cells from the separated suspension wherein these cells or their progeny, or their more matured forms express one or more markers associated with lines of the cellular differentiation. These markers involve: CD14, CD34, CD38, CD45 and ICAM. Hepatic precursor-cells have diameter size 6-16 mc, they are diploid and show indices: glycoforin A-, CD45-, AFP+++, ALB+, ICAM+ and they comprise subpopulations varying with respect to expression of CD14+, CD34++, CD38++ and CD117++. These cells are useful for carrying out cellular and genetic therapy in liver treatment and for preparing artificial organs also.
EFFECT: valuable biological and medicinal properties of cells.
41 cl, 7 tbl, 13 dwg, 15 ex
FIELD: medicine, genetic engineering.
SUBSTANCE: invention relates to applying genetic engineering approaches for treatment of autoimmune diseases, in particular, for treatment of cerebrospinal sclerosis. This is achieved by incorporation of one or some recombinant genes encoding autoantigens that represent a target for autoimmune response. In particular, invention claims a method for designation of gene encoding encephalitogenous epitope of proteolipid protein and expression of gene product in vivo by using the recombinant retroviral vector. Expression and secretion of encephalitogenous epitope improves histopathological and clinical indices in experimental autoimmune encephalomyelitis in mice that is used as a model of cerebrospinal sclerosis. The advantage of invention involves the development of a method for recovery the tolerance in treatment of cerebrospinal sclerosis being without suppression of immune system.
EFFECT: improved and valuable method for treatment.
6 cl, 13 dwg, 3 ex
FIELD: biotechnology and pharmaceutical industry.
SUBSTANCE: title operations are accomplished by following way. Three-dimensional culture of stromal cells is cultured in piston flow bioreactor, in particular being introduced in fibrous matrix incorporated into substrate, which is placed in container constituting a part of bioreactor piston. Stromal cells are grown until density 5 x 106 cell/cm3 substrate is attained, whereupon non-differentiated hemopoietic cells are either sowed directly into piston flow bioreactor or cultured in conditioned medium of stromal cells obtained by gathering medium from indicated flow bioreactor. Non-differentiated hemopoietic cells obtained by cultivation in presence of three-dimensional culture of stromal cells or their conditioned medium are transplanted to into recipient.
EFFECT: enabled growth of large amounts of stromal cells within a relatively small volume to provide longer maintenance of vital activity and reproduction of non-differentiated hemopoietic stem cells or precursor cells.
77 cl, 9 dwg, 3 tbl
FIELD: veterinary science.
SUBSTANCE: the present innovation deals with serological diagnostics of infectious diseases in cattle, viral diarrhea as mucosal disease in cattle (cattle VD) and evaluation of immunity strength in vaccinated animals. The method deals with growing finite cell line of coronary vessels in cow's embryo in Eagle's MEM nutritive medium with 20 mg/ml kanamycin and 10% equine serum. Double dilutions of inactivated tested serumal samples (56 C, 30 min) should be mixed at equal volume (0.05 ml) with 100 TCD50/0.1 ml virus to be incubated for 1 h at 37 C in CO2-incubator. On finishing the incubation one should add 0.1 ml suspension of the above-mentioned finite cell line in Eagle's MEM nutritive medium containing 20 mg/ml kanamycin and 5% (2.5% final concentration) of equine serum. Cell concentration in the culture corresponds to 3.5 x 105. Reaction registration should be carried out in 3 d while cytopathogenic viral action appears in control holes containing working viral dosage. The innovation enables to widen the number of diagnostic immunological methods for veterinary purposes.
EFFECT: higher efficiency of detection.
1 cl, 7 ex, 2 tbl
SUBSTANCE: claimed microarray represents ensemble of gel microcells on substrate made of glass, polymer, ceramic or composite material. Microcells contain immobilized procariotic or eucariotic cells. Microcells with immobilized cells are prepared using gel-forming solution including glycerol. Cellular microarray is used in living cell investigation. In this purpose cellular microarray is incubated in presence of marker, signal (e.g. cell fluorescence) is detected and according to signal level cell living function in microarray is evaluated.
EFFECT: simplified living cell investigation method without losses of cell viability.
12 cl, 5 dwg, 3 ex
FIELD: biology, medicine.
SUBSTANCE: invention relates to media used for reprogramming human and animal stem cells. Medium for the biochemical reprogramming human and animal stem cells containing the medium DI-MEM, fetal calve serum, insulin, 5-azacytidine, L-glutamine comprises additionally the medium F-12, dimethylsulfoxide, dexamethasone, hydrocortisone, N6-2'-dibutyryl-3',5'-cyclic adenosine monophosphate and ethanolamide in the required ratio of components. Invention provides enhancing the effectiveness in reprogramming stem cells in direction of cardiomyogenesis.
EFFECT: improved and valuable properties of medium.
2 dwg, 3 ex
FIELD: agricultural biotechnology, in particular, in vitro grape multiplication processes.
SUBSTANCE: method involves providing micro cutting of testing plants and planting thereof in liquid nutritive medium with reduced amount of macroelements and vitamins, with indole-acetic acid being added in an amount of 0.1-0.3 mg/l and emistim preparation with concentration of 10-7-10-10% being added into composition.
EFFECT: increased efficiency in multiplication of perspective sorts of grape sanitated from virus infection.
FIELD: biotechnology, medicine.
SUBSTANCE: peritoneal macrophages are placed in medium 1999 and subjected for effect of helium plasma obtained at current strength 30 A, voltage 20 V and gas consumption 2 l/min/ Cells are irradiated from distance 20 cm to plasmatron nozzle for 30 s. Method provides reducing time of physical factor effect on cells and allows carrying out the local effect on macrophage functions both in cultured cells and within the whole body. Invention can be used in clinical practice for stimulation of biological processes in cells and tissues.
EFFECT: improved method for stimulation.
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