39 results about "Oil Red O" patented technology

The invention discloses a method for culturing and inducing tilapia mossambica peritoneal preadipocytes. The method comprises the following steps: a) resuspending cells by using a DMEM / F12 culture medium with 10%-15% of fetal calf serum, inoculating the cells into a cell plate, and culturing for 7 days at the temperature of 20-30 DEG C under the condition of 5% CO2; and b) replacing an induction culture medium for sequentially culturing, after inducing for 7-14 days, carrying out oil red O staining of the cells, wherein the fuzzy cell membrane edges can be seen under the microscope, and the interiors of the cells are stained to be red lipid droplets. The invention also discloses a passage method of tilapia mossambica peritoneal preadipocytes. A proliferation culture medium is used for culturing the preadipocytes for 14-20 days, then the preadipocytes are inoculated according to 2 *10<5> to 3*10<5> per milliliter, and the cells can overgrow on the cell plate after the cells are cultured for 2-3 days. The method is capable of successfully carrying out in-vitro culture, induction and passage of the tilapia mossambica peritoneal preadipocytes; the technical reference is provided for the research of the tilapia mossambica lipid metabolism.

The invention provides a method for inducing formation of porcine fat cells by a cell signal channel inhibitor. The method comprises the following steps of uniformly inoculating porcine embryo fibroblasts into a Corning cell culture plate according to a ratio of 4-6* 104 cells / cm2; culturing by using an inducing culture medium, and observing the cells, and performing oil red O dyeing to prove that the cells are fat cells after culturing for 20 days in a culture box containing 5% CO2 at 37 DEG C. The cell signal channel inhibitor instead of a transgene is used for inducing cell transformation,so that genic mutation caused by insertion of an exogenous gene into a cell genome can be effectively prevented; by the method, the research on the forming mechanism of the fat cells is facilitated to a certain extent; and the method with convenience and safety can be widely applied and can be used for researching a fat forming mechanism and also can play a roll in disease therapy and other clinical applications.

The invention belongs to the technical field of biology tissue engineering, and discloses a primary culture and serum-free multidimensional induced differentiation method of adipose-derived stem cells. A collagenase dissociation method is adopted for obtaining the adipose-derived stem cells from subcutaneous and omentum tissue, primary culture is performed until 80% is merged, subculturing is performed, and identification of the fat mesenchyme stem cells is performed. Third generation cells in favorable growth state are selected, a serum-free induced culture medium is added for forming fat andosteogenesis induced differentiation, and finally, oil red O dying and alizarin red dying are performed for identifying the differentiation effect and Real-time RCR is used for detecting the situation of fat formation and osteogenesis gene expression. The adipose-derived cells are easy to obtain, and through primary culture, the fat mesenchyme stem cells are adequate in survival volume and survival period, and ripe lipoblast and osteoblast can be formed through induced differentiation. The primary culture and serum-free multidimensional induced differentiation method provides ideal seed cellsfor cell treatment of metabolic diseases of diabetes, obesity, osteoporosis diseases and the like. A new way is provided for organizational project biolization bones and biolization fat, and the method has wide clinical application prospects.

The invention discloses application of mauremys mutica polypeptide mixtures to the field of preparation of fatty liver prevention and treatment healthcare products or fatty liver prevention and treatment medicines, a fatty liver prevention and treatment healthcare product or a fatty liver prevention and treatment medicine with the mauremys mutica polypeptide mixtures. The application, the fatty liver prevention and treatment healthcare product or the fatty liver prevention and treatment medicine have the advantages that the mauremys mutica polypeptide mixtures have excellent liver healthcare functions; as verified by fatty liver model tests implemented by HepG2 cells induced by different inducers, the contents of TG (triglyceride) and LDL-C (low-density lipoprotein-cholesterol) in the HepG2 fatty liver cells induced by the different inducers including oleic acid, glucose and alcohol can be effectively reduced by the mauremys mutica polypeptide mixtures, the content of lipid in fatty livers of the HepG2 cells induced by the different inducers including the oleic acid, the glucose and the alcohol can be obviously reduced by the mauremys mutica polypeptide mixtures and oil red O staining mauremys mutica polypeptide mixtures which are combined with one another, and accordingly the mauremys mutica polypeptide mixtures have excellent application prospects in the field of preparation of the fatty liver prevention and treatment healthcare products or the fatty liver prevention and treatment medicines.

The invention provides a kit and method for inducing differentiation from rat bone mesenchymal stem cells to fat cells. The kit comprises a fat cell inducing culture solution and a fat cell identification solution, wherein the inducing culture solution comprises a cell culture solution A, a cell culture solution B, a cell culture solution C, a cell culture solution D and a cell culture solution E; the fat cell identification solution comprises a cell immobilizing solution, a coloring agent, a detergent A and a detergent B; the cell culture solution A is an alpha-minimum essential medium (MEM) liquid culture medium; the cell culture solution B is fetal bovine serum; the cell culture solution C is a dexamethasone solution; the cell culture solution D is a 1-methyl-3-isobutylxanthine solution; the cell culture solution E is an insulin solution; the cell immobilizing solution is a paraformaldehyde solution; the coloring agent is an oil red O solution; the detergent A is a 60% isopropanol solution; and the detergent B is double distilled water.

The invention belongs to the technical field of pathological tissue section making, and particularly relates to application of polyethylene glycol to dyeing animal tissue section grease. Through the excellent water solubility characteristic of the polyethylene glycol, with the polyethylene glycol as the dehydrating agent, the permeation and embedding formation of a medium are realized by means ofembedding in the dehydrating process, a formed tissue embedding block has the appearance characteristic of a paraffin block, the embedded medium can be removed by washing the tissue embedding block without processing the tissue embedding block with an organic solvent, and the problem that grease loss is caused by organic solvent processing is avoided. Tissue sections prepared by the method have the thickness of 3-5 micron, and the problem that tissue overlapping is caused by the too high thickness of the frozen sections is avoided. After the tissue sections prepared by the method are dyed by oil red O, due to the dehydration and permeation of the polyethylene glycol, most of grease drops are completely preserved in cytoplasm, the problem of displacement of grease drops in the frozen sections is avoided, and the problem that a tissue structure is not complete since ice crystals are generated in the frozen sections at a low temperature is also avoided.

The invention relates to mesenchymal stem cell differentiation induction liquid and a method for inducing mesenchymal stem cells to adipose cells. The mesenchymal stem cell differentiation induction liquid is prepared from the following components in parts by mass: 0.05mmol / L to 0.2mmol / L of indomethacin, 0.1mmol / L to 0.5mmol / L of 3-isobutyl-1-methylxanthine, 10<-4>mol / L to 10<-6>mol / L of dexamethasone and 5mg / L to 20mg / L of insulin. The mesenchymal stem cell differentiation induction liquid provided by the invention is used for inducing and differentiating the mesenchymal stem cells; after 48 hours, less cytoplasm contains lipid granules with relatively high refractivity can be seen; when culture time is prolonged, the small lipid granules are gathered and gradually become large to form lipid droplets; the quantity of the adipose cells is gradually increased, and round and red lipid droplets with different sizes exist in the mesenchymal stem cells treated by an oil red O dyeing agent.

The invention discloses a method for promoting brown adipose tissue differentiation through SFRP4 and an application. The method comprises treating precursor adipose cells for differentiation by usingSFRP4 active proteins (with different concentrations of 0, 1, 10, 100ng / mL), when the cells grow to 80%, culturing with a cell differentiation induction culture medium I until the cells are fused, adding an induction culture medium II into the cells, inducing for 2-8 days, and replacing the culture medium in the culture bottle with an induction culture medium III, inducing for 2-8 days, observingunder a microscope, after dyeing with oil red O, observing under the microscope again, collecting cells with different differentiation days, extracting total RNA of the cells, detecting expression levels of marker genes UCP-1 and SFRP4 of brown adipose cells by using a real-time quantitative PCR analysis technology after the purity and integrity of the RNA are detected to be qualified, and finally, carrying out statistical analysis. The experimental process is uniform in sampling, and the result reliability is high.

The invention discloses application of a chicken CTGF gene in inhibition of chicken preadipocyte differentiation. The nucleotide sequence of the chicken CTGF gene is as shown in SEQ ID NO.1, and the coded amino acid sequence of the chicken CTGF gene is as shown in SEQ ID NO.2. According to the invention, through a method for adding an exogenous CTGF recombinant protein and constructing an endogenous pCMV-CTGF overexpression vector, an oil red O technology, a Real-time PCR technology and a Western blot technology are utilized to prove that the CTGF gene can inhibit chicken preadipocytes from being differentiated into mature adipocytes from the lipid droplet deposition capability, the transcriptome level and the proteome level of the chicken adipocytes, and thus, the invention provides novel application of the CTGF gene in regulation and control of the chicken preadipocyte differentiation process, and the CTGF gene has practical value in the fields of chicken genetic breeding and animal nutrition.

The invention discloses application of nuclear factor kappa B arrestin 3 combined with A20 in preparation of drugs for treating fatty liver and related diseases, and belongs to new use of ABIN3. An L02 cell line, an ABIN3 over-expressing L02 cell line, a mouse primary hepatocyte cell, and an ABIN3 over-expressing mouse primary hepatocyte cell are used as an experimental object, and a fatty liver cell model is induced through combination of palmitic acid and oleic acid. Oil red O staining results show that compared with L02 cells, an area of red fat droplets in ABIN3 over-expressing L02 cells is significantly reduced under combined stimulation of the palmitic acid and the oleic acid at the same concentration. A same trend of the oil red O staining results is also observed in the ABIN3 over-expressing mouse primary hepatocyte cell, indicating that ABIN3 overexpression significantly inhibits lipid accumulation in fatty liver cells induced by the palmitic acid and the oleic acid. Therefore, ABIN3 has the functions of inhibiting liver lipid accumulation and protecting liver.

The invention discloses a kit for rapid detection of content of triglyceride in fat cells. The kit mainly consists of a Nile red working solution which takes acetone and DMSO as solvents, and PBS. The invention also provides an intracellular triglyceride fluorescent staining method. Compared with a conventional oil red O staining method, an operating method of the kit is simpler, short in time consumption, high in efficiency, high in flux, low in content of impurities and is easier for users to observe, and the operating method is capable of quantifying triglyceride in fat cells within a short time. Therefore, the kit disclosed by the invention is more suitable for current fat cell differentiation researches.

The invention relates to the technical field of stem cell culture, in particular to a culture medium, application thereof and a method for inducing differentiation of tendon stem cells into adipocytes. TGF-beta1, EGF and G-CSF are added to a basal medium so as to obtain an induction culture liquid for differentiation of the tendon stem cells into adipocytes, the number of the tendon stem cells which are differentiated into adipocytes is increased, and the time of differentiation is shortened. It is shown through experiments that the tendon stem cells which are induced by the culture liquid canexhibit the characteristics of adipocytes within 21 days, the cell morphology is changed, and the number of Oil red O stained cells is increased; and after 28 days of induction, the cell morphology has gradual change and a polygonal shape, calcified nodules formed through mineral deposition are contained in intercellular substances, and the staining characteristic of oil red O is positive.

The invention discloses a method for efficiently separating intramuscular fibroblast-lipogenic progenitor cells of a mouse. The method comprises the following steps that (1), a collagen coating culture dish is prepared; (2), mouse skeletal muscle is separated and digested; and (3), differential adherence separation is carried out on the FAP. It is determined through cell flow analysis and oil red O staining that the separated cells are high in purity, high in differentiation potential, high in proliferation speed and high in survival rate; by adopting the collagen coating culture dish, cell attachment is facilitated, the separation time is greatly shortened, and the cell yield is improved and the used C57BL / 6 mouse is wide in source and easy to obtain, and the difficulty of reproduction of an experimental method is greatly reduced. Compared with a traditional flow sorting method, a flow sorter is not needed, and the method is more economical, practical, convenient and rapid; and a good cell basis is provided for researching the functions of the fibroblast / lipoblastic progenitor cells in development and regeneration of skeletal muscles.

The invention discloses application of a Bromodomain-containing protein 4 (BRD4) and an inhibitor thereof in preparing drugs for treating fatty liver and related diseases. Different liver cells (L02 cell and HepG2 cell) are adopted as experimental subjects which are treated by BRD4 inhibitors, JQ1 AND OTX015 respectively; with DMSO as control, the liver cells are treated by palmitate / oleic acid (PA / OA), and then are dyed by oil red O; the dyeing results show that: in comparison with the DMSO control group, the color of the inhibitor treatment group obviously becomes light, indicating that after BRD4 is inhibited, fat accumulation can be suppressed, and the occurrence of fatty liver and related diseases can be reduced. Therefore, the BRD4 provides a target of developing drugs for preventing, relieving and / or treating fatty liver and related diseases.

The invention belongs to a novel application of paeonol, and particularly relates to an application of paeonol in preparation of medicines for treating obesity and lipotoxic cardiomyopathy. In a model of treating mouse obesity by paeonol, through animal characterization and oil red o staining of rat primary myocardial cells, the effect of paeonol in glycolipid metabolism change and heart lipid metabolism disorder caused in a mouse obesity model fed with high fat diet is researched. The results show that paeonol can alleviate glycolipid metabolism change and heart lipid toxicity caused by obesity induced by high fat diet. Paeonol may become a potential effective treatment method for heart lipotoxicity caused by obesity.

The invention discloses application of a nuclear factor chiB inhibition protein 1 combined with A20 to preparation of a medicine for treating fatty livers and related diseases and belongs to novel application of ABIN1 (A20binding Inhibitor of NF-KB Activation). According to the application, an L02 cell line, an ABIN1 over-expression L02 cell line, mouse primary hepatocyte cells and ABIN1 over-expression mouse primary hepatocyte cells are adopted as experiment targets, fatty liver cell models are induced through palmitic acid and oleic acid together, and oil red O dyeing results show that undercombined irritation of the palmitic acid and the oleic acid of a same concentration, the areas of red fat granules in ABIN1 over-expression L02 cells are remarkably reduced when being compared with those of L02 cells. The same oil red O dyeing result trend is also observed in mouse primary hepatocyte cells with over-expressed ABIN1, the result shows that ABIN1 over-expression has a remarkable inhibition function on lipid accumulation in palmitic acid and oleic acid induced fatty liver cells. Therefore, ABIN1 has functions of inhibiting liver lipid accumulation and protecting livers.

The invention provides a method for inducing formation of porcine fat cells by a cell signal channel inhibitor. The method comprises the following steps of uniformly inoculating porcine embryo fibroblasts into a Corning cell culture plate according to a ratio of 4-6* 104 cells / cm2; culturing by using an inducing culture medium, and observing the cells, and performing oil red O dyeing to prove that the cells are fat cells after culturing for 20 days in a culture box containing 5% CO2 at 37 DEG C. The cell signal channel inhibitor instead of a transgene is used for inducing cell transformation,so that genic mutation caused by insertion of an exogenous gene into a cell genome can be effectively prevented; by the method, the research on the forming mechanism of the fat cells is facilitated to a certain extent; and the method with convenience and safety can be widely applied and can be used for researching a fat forming mechanism and also can play a roll in disease therapy and other clinical applications.

The invention belongs to the technical field of biological medicine, and relates to a naphthalene-indandione donor-acceptor compound, a preparation method thereof and application of the naphthalene-indandione donor-acceptor compound in a lipid droplet no-wash fluorescent probe. The chemical structure is shown as a formula I, wherein R1 and R2 are respectively and independently hydrogen, lower alkyl, ether group, substituted alkyl or acyl; r3, R4, R5 and R6 are respectively and independently hydrogen, methyl, fluorine, chlorine, bromine, iodine, cyano, trihalomethyl, nitryl, phenyl or pyridyl. When the naphthalene-indandione donor-acceptor compound provided by the invention is used as a lipid droplet fluorescent probe, very weak fluorescence is shown in a high-polarity solvent, but fluorescence emission is remarkably increased in a low-polarity environment, so that lipid droplets can be lighted in a washing-free manner with high selectivity and high contrast; meanwhile, selective distinguishing of fatty liver tissues can be successfully realized. Compared with the existing oil red O staining, the fluorescent probe has the advantages of high sensitivity, good selectivity, time saving, simplicity in operation, low incubation concentration and the like.

The invention belongs to the technical field of cell model construction, and discloses a metabolism-related fatty liver disease in-vitro cell model and a construction method thereof.HepG2 and LO2 cells are treated with OA or PA or a mixture thereof respectively, and the cell survival rate is detected with CCK8; detecting the lipid accumulation degree of the cells by oil red O staining and a triglyceride enzyme method; according to the present invention, qRT-PCR is adopted to detect the expression levels of apoptosis-related proteins Bax, Bcl2 and Cleved caspase-3, fibrosis-related proteins alpha-SMA and Col.I, autophagy-related proteins LC3-II, P62 / SQSTM1 and Beclin-1, and inflammatory factors NLRP3 and TNF-alpha, the saturated fatty acid PA is used for treating HepG2 cells, so that a certain degree of lipid accumulation can be caused, remarkable cell damage and fibrosis are induced, and the MAFLD in-vitro cell model is a proper MAFLD in-vitro cell model.

The invention provides an application for a crude broussonetia papyrifera leaf extract in preparation of drugs used for treating the non-alcoholic fatty liver disease. Through measurement of the contents of triglyceride and total cholesterol in cell oil red O staining, mouse liver oil red O staining and mouse blood, the crude broussonetia papyrifera leaf extract is proved to be able to effectivelyinhibit aggregation of lipid in HepG2 cells, improve lipid accumulation in the liver and reduce the level of the cholesterol and the triglyceride in the mouse blood. According to the invention, in atest of treating the non-alcoholic fatty liver disease with the crude broussonetia papyrifera leaf extract, no obvious toxic effect is shown; meanwhile, the crude broussonetia papyrifera leaf extracthas good curative effect. Thus, the crude broussonetia papyrifera leaf extract can be used for development of new drugs, and has important significance on drug target confirmation.

The invention discloses application of DHC (dehydrocorydaline) in preparation of a preparation for treating atherosclerosis. ApoE<- / ->mice prove that formation of atherosclerotic plaques treated by the DHC is reduced, the lesion area of the plaques is reduced, and it is indicated that DHC can relieve development of atherosclerosis. An oil red O staining result shows that the grease area of an administration group is reduced, F4 / 80 immunofluorescence staining shows that macrophage infiltration of the administration group is less, and a Masson staining result shows that the fiber collagen content of the administration group is higher, which indicates that the DHC can improve the stability of plaques. Administration of the DHC can also reduce systemic inflammation of atherosclerosis. An important scientific evidence is provided for potential application of the DHC as an atherosclerosis treatment or improvement agent.

The invention provides a method for quantitatively detecting the deposition amount of mesenteric fat in fish and application. Based on the characteristic that fat-soluble dye oil red O can specifically color adipose tissues, the method for accurately and conveniently quantifying the deposition of mesenteric fat in fish is developed. A dyeing-extraction technical scheme is adopted, the problems of rough quantification, large errors, inconvenience in operation and the like of a traditional method for directly weighing adipose tissues scraped by a scalpel are solved, and the deposition amount of the mesenteric fat of an individual is obtained through the operation steps of visceral mass sample collection and treatment, sample marking, fixing, dehydration, constant dyeing, rinsing, extraction, quantification and the like. the deposition amount is expressed by the mass of oil red O adsorbed and extracted by a sample, and the method can be used for accurately comparing the difference of mesenteric fat deposition among fish individuals.

The invention discloses application of OTUB2 and an OTUB2 inhibitor to preparing a medicine for treating fatty liver and related diseases. An LO2 cell is taken as the experimental subject to respectively construct OTUB2 overexpressed and OTUB2 knock-down LO2 cell models, and oil red O staining is conducted after the simulation by palmitic acid / oleic acid (PA / OA). The staining result shows that: compared with a phage control group, the staining of an OTUB2 overexpression group is deepened, and the staining of an OTUB2 knock-down group becomes shallow. The result shows that the fat accumulationis promoted and the fatty liver is aggravated after the overexpression of an OTUB2 gene; the fat accumulation is inhibited after the OTUB2 gene is knocked out, and the occurrence of the fatty liver isimproved. Therefore, the OTUB2 provides a target for the preparation of the medicine used for preventing, relieving and / or treating the fatty liver and the related diseases.

The invention discloses application of mauremys mutica polypeptide mixtures to the field of preparation of fatty liver prevention and treatment healthcare products or fatty liver prevention and treatment medicines, a fatty liver prevention and treatment healthcare product or a fatty liver prevention and treatment medicine with the mauremys mutica polypeptide mixtures. The application, the fatty liver prevention and treatment healthcare product or the fatty liver prevention and treatment medicine have the advantages that the mauremys mutica polypeptide mixtures have excellent liver healthcare functions; as verified by fatty liver model tests implemented by HepG2 cells induced by different inducers, the contents of TG (triglyceride) and LDL-C (low-density lipoprotein-cholesterol) in the HepG2 fatty liver cells induced by the different inducers including oleic acid, glucose and alcohol can be effectively reduced by the mauremys mutica polypeptide mixtures, the content of lipid in fatty livers of the HepG2 cells induced by the different inducers including the oleic acid, the glucose and the alcohol can be obviously reduced by the mauremys mutica polypeptide mixtures and oil red O staining mauremys mutica polypeptide mixtures which are combined with one another, and accordingly the mauremys mutica polypeptide mixtures have excellent application prospects in the field of preparation of the fatty liver prevention and treatment healthcare products or the fatty liver prevention and treatment medicines.

The invention discloses an application of a PMX-53 polypeptide in preparation of a medicine for treating non-alcoholic steatohepatitis. After the PMX-53 inhibitor is used for treatment, Hamp is converted from Hamp to Hamp; e and oil red O staining patterns show that the number of lipid droplets in hepatocytes of NASH mice is reduced, the lipid droplets become small, and liver fat is obviously reduced (Figure 3A); quantitative PCR results show that expression of fat synthesis related genes Srebf1, Acc and Fasn is remarkably reduced (figure 3B), WB is further used for confirmation, and figure 3C results show that fat synthesis related proteins Acc and Fasn in mouse liver tissues are reduced; quantitative PCR (Polymerase Chain Reaction) and ELISA (Enzyme-Linked Immunosorbent Assay) detection show that the expression of the inflammation-related factors IL-6 and TNF-alpha is obviously reduced ( The results show that the non-alcoholic steatohepatitis of mice can be relieved by treating with the C5ar1 specific inhibitor PMX-53, so that the PMX-53 can be applied to preparation of the medicine for treating the non-alcoholic steatohepatitis.

The invention discloses an application of an ITGA2 gene in regulating and controlling the formation of rice-colored fat of a pig. According to the present invention, the RNA-seq technology is adopted, the gene expression difference of the mature beige fat cells and the white fat cells of the Tibetan pig is analyzed, the beige fat specific high expression gene is screened, and the pig specific beige fat regulation gene ITGA2 is successfully identified based on the human, mouse and pig comparative genomics technology; the expression of the ITGA2 gene is knocked down by further utilizing an RNAi technology, and it is proved through oil red O dyeing and a seharse experiment that ITGA2 is needed for the formation and function maintenance of the rice-colored fat of the pig. According to the invention, the effect of ITGA2 in the formation process of the rice-colored fat of the pig is discovered for the first time, and the discovery has important significance for regulating and controlling fat deposition of the pig by utilizing the rice-colored fat.