50 results about "Noggin" patented technology

The invention discloses a method for building medicine transport models for research on BCRP (breast cancer resistance proteins) mediation for 3D (three-dimensional) organs of the small intestines of mice. The method includes separating crypts from the small intestines of the mice by means of digestion, suspending the crypts in matrigel and then joining the crypts with cell culture plates and promoting differentiation of the crypts by the aid of ADMEM/F12 media with Respondin-1, m-noggin and m-EGF cell differentiation growth factors to form the 3D organs; carrying out morphologic observation and detecting the expression level of BCRP genes and proteins to verify the feasibility of model theories; carrying out research on the trans-membrane transport activity of BCRP in the 3D organs by the aid of fluorescent substrates Hoechst 33342 of the BCRP and inhibitors Ko143 or YHO-13177 of the fluorescent substrates by co-incubation processes. The method has the advantages that the medicine trans-cell-membrane transport in-vitro models for the research on the BCRP mediation for the 3D organs of the small intestines of the mice are built for the first time, and the method for building the models is easy and convenient to implement and high in detection efficiency and speed and can be widely applied to screening BCRP substrates and inhibitors in an in-vitro manner.

The invention puts forward a universal cancer organoid in-vitro culture medium, which consists of a DMEM/F-12K (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12K) basic culture medium and an adding factor, wherein the adding factor consists of HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), L-Glutamine, EGF (epidermal growth factor), Noggin, FGF (fibroblast growth factor)-10, A83-01 and Y27632. According to the culture medium of the embodiment of the invention, the success rate of organoid passage culture can be greatly improved, in addition, the universal culture of cancer samples, including gastric carcinoma, rectal carcinoma, lung carcinoma and the like, can be realized, culture cost is greatly lowered, the long-term culture of the organoid can be realized, and abiobank is established.

The invention relates to the fields of cytobiology and cell culture, and particularly relates to a breast cancer organoid culture kit. A method for realizing the kit comprises the following steps: (1)preparing, sub-packaging and using a breast cancer organoid culture medium A; (2) preparing, sub-packaging and using a breast cancer organoid culture medium B; and (3) preparing, sub-packaging and using a sample preserving fluid. The breast cancer organoid culture medium A is a DMEM/F12 culture medium containing 100 U/ml of penicillin, 0.1 mg/ml of streptomycin, 1% of HEPES and 1% of Gluta Max 100 X; the organoid culture medium B is a DMEM/F12 culture medium containing EGF, an FGF growth factor, an ALK inhibitor A83-01, Noggin, an HEPES buffer solution, Y27632 and one or more of a Wnt signalpath related secretory protein family R-Spondin 1-4; and the sample preserving fluid is a DMEM/F12 culture medium containing 100 U/ml of penicillin, 0.1 mg/ml of streptomycin and 1% of L-glutamine. The three components form the breast cancer organoid culture kit.

The present invention provides a culture medium and a culture method for colorectal cancer organoids. The culture medium comprises a basic culture medium Advanced DMEM/F12, specific additive factors and sterile water; wherein a mass ratio of the basic culture medium Advanced DMEM/F12 to the sterile water is 99:1; and the specific additive factors comprise B27 without vitamin A, N-acetylcysteine, EGF, Noggin, R-spondin 1, Wnt3a, CHIR99021, thiazovivin, Gastrin I, a penicillin-streptomycin mixture and Primocin. The culture medium is used to culture the colorectal cancer organoids, can maintain morphological structures and genetic characteristics of primary tissues, effectively reduces risks of microbial contamination in colorectal cancer culture, and improves success rate and survival rate of colorectal cancer organoid culture.

A method for generating chondrocytes and/or cartilage, optionally articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue and/or hypertrophic chondrocyte like cells and/or cartilage like tissue, the method comprising: a. culturing a primitive streak-like mesoderm population, optionally a CD56+, PDGFR[alpha]+KDR- primitive streak-like mesoderm population, with a paraxial mesoderm specifying cocktail comprising: i. a FGF agonist; ii. a BMP inhibitor; optionally Noggin, LDN-193189, Dorsomorphin; and iii. optionally one or more of a TGF[beta] inhibitor, optionally SB431524; and a Wnt inhibitor, optionally DKK1, IWP2, or XAV939; to specify a paraxial mesoderm population expressing cell surface CD73, CD105 and/or PDGFR-beta; b. generating a chondrocyte precursor population comprising: i. culturing the paraxial mesoderm population expressing CD73, CD105 and/or PDGFR-beta at a high cell density optionally in serum free or serum containing media; ii. culturing the high cell density CD73+, CD105+ and/or PDGFR[beta]+ paraxial mesoderm population with a TGF[beta]3 agonist in serum free media to produce a high cell density Sox9+, collagen 2+ chondrocyte precursor population; and c. either i. culturing the high cell density Sox9+, collagen 2+ chondrocyte precursor population with the TGFbeta3 agonist for an extended period of time to produce an articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue; or ii. culturing the high cell density Sox9+ collagen2+ chondrocyte precursor population with a BMP4 agonist for an extended period of time to produce a hypertrophic chondrocyte like cells and/or cartilage like tissue.

The invention discloses a culture medium and a culture method for constructing a liver tumor scaffoldless organoid. The culture medium comprises the following components: Advanced DMEM/F12, FBS, diabody, N-2, Noggin, B-27, EGF, FGF-10, Y-27632, A83-01, SB202190, R-Spondin, N-acetylcysteine, Nicotinamide, Wnt3A, HGF and PrimocinTM. The culture method comprises the steps of adding primary tumor cells separated and obtained into the above-mentioned special culture medium for resuspension, inoculating into an ultra-low adsorption U-shaped well plate after counting, and centrifugally culturing until organoids are formed. The culture medium and the culture method can realize in-vitro proliferation of primary liver cancer cells, can quickly construct the liver tumor scaffoldless organoids, and can maintain long-term stable growth of the liver tumor organoids. The tumor organoids formed by the culture medium and the culture method retain heterogeneity of tumor tissues of patients, and are suitable for drug sensitivity detection, tumor generation and development related research of in-vitro liver cancer chemotherapeutic drugs.

The invention discloses a method for inducing human embryonic stem cells to differentiate to neuroderm. The method comprises the following steps that H9 cells are kept to passage in a cell cluster state, wherein the cell clusters are uniform in size, and high-density culture of the cell clusters can be kept; based on this, two inducing factors of SB431542 and NOGGIN are introduced, a knock out serumreplacer (KSR) culture medium and an N2 culture medium which are determined according to chemical components are used in matched manner according to different proportions at different culture times; and ascorbic acid (vitamin C) is added into the N2 culture medium at induced differentiation later period, so that HESC can be induced within shorter time to differentiate directionally to form an NE cell for expressing a PAX6 protein.

The invention discloses a standardized culture medium and a culture method for three-dimensional culture of lung and lung cancer tissue organoids. The standardized culture medium comprises at least one or more of a reagent A and a reagent B, wherein the reagent A comprises one or more of R-spondin1, Noggin, N-acetylcysteine, nicotinamide, Y-27632, A8301, SB202190, FGF-7 and FGF-10; the reagent B comprises a reagent B1, a reagent B2 and a reagent B3; the reagent B1 comprises one or more of bovine pituitary protein extract, Y-27632, DAPT, hydrocortisone, insulin, gentamicin, amphotericin, tretinoin, transferrin, iodomethacin, epinephrine and human epidermal growth factors; the reagent B2 comprises heparin; and the reagent B3 comprises one or more of DMEM/F12, L-ascorboic acid-2-phosphate magnesium and sodium selenium. According to the standardized culture medium and the culture method, aiming at the growth characteristics of adult stem cells in lung tissues, multiple growth factor components are selected for blending, and by optimizing the proportion of growth factors in the culture medium, normal lung tissues and lung cancer cells can effectively form lung and lung cancer organoidsin a three-dimensional culture environment.

The invention discloses a culture medium special for organoids of nasopharyngeal carcinoma and a scaffold-free culture method. The culture medium comprises EGF, Noggin, Y-27632, A83-01, SB202190, bFGF, hydrocortisone, Insulin, penicillin-streptomycin, FBS and Keratinocyte-SFM. The culture method comprises the following steps: inoculating separated tumor cells of nasopharyngeal carcinoma into an ultra-low adsorption U-shaped cell culture plate, adding the culture medium special for organoids of nasopharyngeal carcinoma, performing centrifugation, performing culture in a CO2 incubator with a volume concentration of 5% at a temperature of 37 DEG C, and replacing a scaffold-free culture medium once every 3-4 days. According to the culture medium special for organoids of nasopharyngeal carcinoma and the scaffold-free culture method, introduction of an exogenous scaffold material is not needed, cell aggregation and clustering can be promoted under the assistance of gravity, rapid amplification of the tumor cells in a short time is achieved so as to construct the organoids, the success rate can reach 100%, and the specificity of primary tumor can be maintained; and meanwhile, the activityand functional and high expression of the tumor cells can be maintained for a long period of time.

The invention relates to a biomedical technology, in particular to a culture medium and culture method for ovarian cancer organoids. The culture medium comprises a basal culture medium DMEM/F12, B27, glutamine, HEPES, N-acetylcysteine, FGF2, A83-01, nicotinamide, forskolin, beta-estradiol, hydrocortisone, R-spondin-1, noggin, FGF10, EGF and Heregulin beta 1 and at least one selected from PFBS, PFOA and PFOS. According to the culture medium and the culture method, the establishment success rate of an ovarian cancer culture medium is improved, the amplification efficiency of the ovarian cancer organoids is improved, the proliferation efficiency after passage is stable, the drug screening process is accelerated, and the cost of culture materials is reduced.

The invention discloses a standardized culture medium for three-dimensional culture of intestinal and intestinal cancer tissue organs and a culture method. The standardized culture medium comprises atleast one or more of a reagent A, a reagent B and a reagent C; wherein the reagent A comprises one or more of Nacetylcysteine, Y-27632, A8301, a recombinant human epidermal growth factor and gastrin;and the reagent B is prepared from one or more of R-spodin1, Noggin, nicotinamide and SB202190, and the reagent C comprises Wnt-3A. According to the culture medium disclosed by the invention, aimingat the growth characteristics of adult stem cells in intestinal tissues, various growth factor components are selected for blending, and by optimizing the proportion of growth factors in the culture medium, the adult stem cells of intestines and cells of intestinal cancer can effectively form organoids of intestines and organoids of intestinal cancer respectively in a three-dimensional culture environment.

The invention discloses application of BMP2 and Noggin in jointly preparing an ankylosing spondylitis diagnostic reagent kit.The invention further discloses application of BMP2 and Noggin in jointly preparing a reagent kit for predicting the severity of ankylosing spondylitis pathologic osteogenesis.The invention further discloses a BMP2 detection reagent kit and a Noggin detection reagent kit.By detecting the BMP2 level and the Noggin level at the same time, the specificity and sensitivity of diagnosis of ankylosing spondylitis can be improved, and the severity of ankylosing spondylitis pathologic osteogenesis can be predicted.By means of the BMP2 detection reagent kit and the Noggin detection reagent kit, the lower limit of the reagent kit detection allowable range is lowered, and the detection accuracy is improved.

The invention provides a culture medium of endometrial organ and a culture method. The culture medium is prepared from the following components according to final concentration: EGF with the concentration of 10 to 100 ng/ml, Noggin with the concentration of 20 to 500 ng/ml, R-spondin 1 with the concentration of 20 to 500 ng/ml, Wnt3a with the concentration of 20 to 500 ng/ml, FGF9 with the concentration of 1 to 100 ng/ml, KIAA1199 with the concentration of 10 to 200 ng/ml, sox2 with the concentration of 1 to 100 ng/ml, isoflavone with the concentration of 10 to 100 nM, CHIR99021 with the concentration of 100 to 2000 nM, A83-01 with the concentration of 1 to 40 [mu] M, a ROCK inhibitor with the concentration of 2 to 50 [mu] M and primary cell antibiotic with the concentration of 50-200 [mu]/ml; and all the components are dissolved in a DMEM/F12 culture solution. When the culture medium is used for culturing endometrial organ, rapid amplification of endometrial stem cells and maturation and differentiation of the organ are facilitated, the number of passage times is remarkably increased, and the same tissue structure and genome characteristics of primary tissues can still be maintained after multiple passage times.

The invention discloses a method for qualitative identification of strawberry stigma protein based on the two-dimensional liquid chromatography tandem mass spectrometry platform, comprising the following steps: 1) carrying out extraction and purification of strawberry stigma to obtain protein powder; 2) carrying out protein quantification and reductive alkylation; 3) carrying out proteolysis; 4) carrying out first-dimensional SCX strong cation exchange liquid chromatography separation of a peptide mixture; 5) carrying out second-dimensional reverse C18 liquid phase separation tandem Q Exactivemass spectrometry; and 6) carrying out data analysis: The original files obtained by the mass spectrometer were identified by the Maxquant search library to obtain a protein and peptide information list. The method of the invention can be adopted for qualitative identification of the red-cheek strawberry stigma protein, and provides a basis for building the red-cheek strawberry stigma protein information database.

The invention discovers that a Noggin gene generates a regulation and control effect on the villus fineness of Liaoning cashmere goat, that is to say, the higher expression quantity of the Noggin gene can promote the expression quantity of KAP7.1 and KAP8.2 genes to be increased, the increase of the genes can reduce the villus fineness and improve the villus quality, and thus an application of the Noggin gene as an exogenous gene introduced into cashmere goat cells to improve the villus fineness is disclosed; Noggin gene over-expressed lentivirus is injected into a male Liaoning cashmere goat testis, and after the KAP26.1 gene as an exogenous gene is introduced into the cashmere goat cells through virus injection, a Liaoning cashmere goat individual having high expression quantity of the KAP7.1 and KAP8.2 genes is selected and bred as a breeding sheep, so that the villus fineness of the cashmere goat can be improved, and the villus quality is increased.

The present invention relates to a pharmaceutical composition or a cosmetic composition for preventing or treating hair loss, or promoting hair growth. The composition according to the present invention exhibits an excellent effect of preventing or treating hair loss and promoting hair growth by comprising cyclic ADP ribose (cADPR) or derivatives thereof or comprising at least one selected from the group comprising one or more nature-derived amino acid or salt thereof, one or more growth factor, noggin, one or more saturated or unsaturated C8 to C18 long chain fatty acid or salt thereof, one or more active factor and one or more water-soluble vitamin or salt thereof in addition to cyclic ADP ribose, and can be safely used regardless of sex and age.

The invention provides a composition, a culture medium and a method for 3D culture of laryngeal cancer tissues. The culture medium comprises the following components according to final concentration: 0.5-2 * of a B27 serum-free additive; the concentration of Wnt3A is 50 to 500 ng/ml; the concentration of the N-acetylsteine is 0.15 to 1.5 mM; the concentration of the EGF is 20 to 100 ng/mL; the concentration of Noggin is 50 to 200 ng/mL; the concentration of the R-spondin 1 is 200 to 1000 ng/mL; the molecular weight of GSK1838705A is 0.2 nM to 2 nM; the concentration of the FGF10 is 10 to 50 ng/mL; 5 to 20 mM of Nicotinamide, and 10 to 20 mM of Nacotinamide; y-27632, 1 to 20 [mu] M; the solvent is an SAGM culture medium. According to the culture medium for the laryngeal cancer tissue organoid, disclosed by the invention, aiming at the culture and growth characteristics of laryngeal cancer tissue source cells, various cell factor components are selected and prepared according to a certain proportion, and laryngeal cancer cells can effectively form the organoid in the 3D culture medium; the formed organoid can maintain high consistency of tissue cell specificity, stem cell characteristics and genetic typing and high similarity of cellular morphology and physiological functions.