111 results about "In vitro growth" patented technology

Compositions and methods for the growth and expansion of mammalian cells in culture are provided. In particular, methods for the growth and expansion of postpartum-derived cells in vitro are provided using surfaces such as microcarrier beads.

The present invention provides adipose-derived stem cells and lattices. In one aspect, the present invention provides a lipo-derived stem cell substantially free of adipocytes and red blood cells and clonal populations of connective tissue stem cells. The invention also provides a method of isolating stem cells from adipose tissues. The cells can be employed, alone or within biologically-compatible compositions, to generate differentiated tissues and structures, both in vivo and in vitro. Additionally, the cells can be expanded and cultured to produce hormones and to provide conditioned culture media for supporting the growth and expansion of other cell populations. In another aspect, the present invention provides a lipo-derived lattice substantially devoid of cells, which includes extracellular matrix material from adipose tissue. The lattice can be used as a substrate to facilitate the growth and differentiation of cells, whether in vivo or in vitro, into anlagen or even mature tissues or structures.

Methods for the maximizing parameter of the in vitro growth and expansion of mammalian cells, specifically postpartum-derived cells in containers such as roller bottles is described. Methods of optimizing growth rate and cell yield in such culture systems are provided. The methods are particularly adapted for human postpartum-derived cells, such as umbilicus-derived cells.

The present invention discloses a method for improving growth and survival of single human embryonic stem cells. The method includes the step of obtaining a single undifferentiated HES cell; mixing the single undifferentiated cell with an extracellular matrix (ECM) to encompass the cell; and inoculating the mixture onto feeder cells with a nutrient medium in a growth environment. Therefore the single cells can survive, proliferate and grow in vitro.

The invention provides a preparation method for a chlorella polypeptide microcapsule. The method comprises the following steps: extracting chlorella protein by using a low temperature ultrahigh pressure continuous flow cell crusher; hydrolyzing the chlorella protein with papain; filtering the hydrolyzed chlorella protein with an ultrafiltration centrifuge tube; then carrying out separation and purification by using ion exchange chromatography DEAE-52 and gel chromatography dextrangel G-25 columns; and finally utilizing complex coacervation to prepare the chlorella polypeptide microcapsule. The chlorella polypeptide microcapsule prepared in the invention has antitumor activity, e.g., the chlorella polypeptide microcapsule has inhibitory effects on in vitro growth of human hepatoma carcinoma cells HepG2 and the inhibition rate of the chlorella polypeptide microcapsule reaches 38% when concentration is 400 mu g / mL. Therefore, the chlorella polypeptide microcapsule prepared in the invention is beneficial for development and utilization of antitumor health food and medicinal products.

The invention relates to an erythroculter ilishaeformis spermatogonia stem cell separation and culture method. The method comprises the following steps: (1) collecting spermatic tissues: collecting spermary of erythroculter ilishaeformis with an age of seven months in a sterile environment, then rinsing spermary in PBS containing double-antibody, and finally grinding the spermary to disperse the spermary tissues; (2) digesting and separating spermatogonia stem cell: adding IV type collagenase (0.1%) into the dispersed spermary tissues, subjecting the digestive fluid to centrifugal separation, collecting the precipitate, digesting the precipitate by trypsin (0.25%), stopping the digestion by a culture medium containing FBS (10%), and collecting the cells; (3) carrying out primary culture of spermatogonia stem cell: using a DMEM / F12 complete medium containing cell factors to re-suspend the cells obtained in the step (2), adjusting the cell concentration, then paving the suspension liquid on a 24-hole cell culture plate coated by gelatin, and culturing the cells at a temperature of 26 DEG C. Trough the provided method, the in-vitro growth of primary cells of erythroculter ilishaeformis spermatogonia stem cell becomes easier, and an effective cell platform is provided for the research on reproduction and growth of erythroculter ilishaeformis.

The present invention describes method for culturing preadipocytes isolated ex vivo, the method including introducing preadipocytes into a three dimensional support matrix, and allowing the cells to differentiate in vitro into adipocytes within the support matrix. The matrix may be a collagen matrix. The method may be used for investigating the development of stem cells, or for investigating the response of adipocytes to stimuli. The method provides a system whereby adipocytes with biological properties resembling those in vivo can be grown in vitro.

The invention provides a cell culture process and a method for the in vitro growth of microvascular endothelial cells, including myometrial cells and diagnostic, therapeutic and prophylactic applications of microvascular endothelial cells.

The invention discloses a method and a system for carrying out magnetic micromanipulation on a cell in a physiological environment. The system for carrying out the magnetic micromanipulation on the cell in the physiological environment comprises a magnetic probe, a probe nanometer offset detection module, a three-dimensional displacement platform, an inverted / upright optical microscope, a magnetic probe control module and a displacement control module and realizes the in-vitro operation and real-time tracking observation of a biological cell by being placed into a fine controlled culture environment suitable for the in-vitro growth of the cell. The method for carrying out the magnetic micromanipulation on the cell is characterized by measuring a single cell which contains magnetic nanometer particles point by point according to a selected area by utilizing the magnetic probe on the basis that the magnetic micromanipulation on the cell is combined with the magnetic nanometer particles, recording the magnitude of magnetic force, simultaneously acquiring the magnetic force and shape dot images at a nanometer scale and operating, moving, carrying, injecting and testing the single cell by using the magnetic probe by accurately positioning the magnetic cell and the magnetic nanometer particles, thereby achieving the guiding significance on a nanometer biological technology and micro-nanomanipulation technologies.

The invention relates to a system and method for measuring electrical characteristics of biological cells through a nano-electrode array under physiological conditions. The system consists of the nano-electrode array, an inverted or upright optical microscope, a probe control module and an electrical signal processing module. The system is arranged in a culture environment suitable for cell in-vitro growth, the electrical characteristics of a single cell are detected through the nano-electrode array and probes, the strength of an electrical signal is measured and recorded according to selected points or areas, and the electrical characteristics of a cell body are obtained on a nano-scale. The invention aims to improve an existing micro-electrode-array-based method for measuring the electrification amount of the biological cells by using the nano-electrode array so as to provide the method and system capable of measuring the electrification amount of the biological cells, applying an electric excitation signal to the single cell for observing the change of the single cell and also detecting the electrical characteristics of the cell by utilizing the probes.

The present invention discloses the use of one kind of tremella heteropolysaccharide or its extract in preparing medicine, beverage, food and health product for promoting the growth and propagation of probiotics, in preparing medicine, beverage, food and health product for treating constipation, in preparing medicine, beverage, food and health product for expelling toxin and nursing face, and in preparing promoter for promoting the in vitro growth and propagation of probiotics.

The invention is directed to a chemically defined animal cell culture media, and methods for preparing such a medium, wherein the media are suitable for culturing epidermal cells, preferably human epidermal cells, including cells of the hair follicle. The invention further provides for methods of culturing epidermal cells, hair follicles, and skin explants in the media as well as uses of the cell cultures and explant cultures in screening assays.

The invention is directed to a chemically defined animal cell culture media, and methods for preparing such a medium, wherein the media are suitable for culturing epidermal cells, preferably human epidermal cells, including cells of the hair follicle. The invention further provides for methods of culturing epidermal cells, hair follicles, and skin explants in the media as well as uses of the cell cultures and explant cultures in screening assays.

The invention discloses a novel BH3 analogue targeted to Bcl-2 family anti-apoptotic protein and application of the novel BH3 analogue. The structural general formula of the BH3 analogue is as shown in the general formula A (the general formula A can be found in specification). A thought and method of peptidomimetics is adopted, the structure of a natural BH3 peptide fragment is simplified or modified, and the novel BH3 analogue is obtained. It is proved by experiments that the compound as shown in the general formula A and the Bcl-2 family anti-apoptotic protein represent excellent binding activity on the aspect of the molecular level; and it is shown by in-vitro carcinoma cell growth inhibition experiments that the compound has a certain in-vitro growth inhibition function on the human chronic myeloid leukemia cell K562, the human promyelocytic lenukemia cell HL-60 and the human tissue cell lymphoma cell U937. It is prompted by research results that the compound can be used as a candidate drug for preventing or treating related diseases caused by expression abnormality of anti-apoptotic protein in the Bcl-2 family protein.

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated / progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and / or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth / regeneration.

The invention relates to a method for constructing animal models, in particular to a method for constructing oral mucosa carcinoma animal models. The method adopts the technical proposal that the method for constructing the oral mucosa carcinoma animal models comprises the following steps: firstly, culturing a tumor tissue and tumor cells; and identifying biological characteristics of in vitro growth of the cells. The method for constructing the oral mucosa carcinoma animal models has the advantages that: firstly, Rca-B cell lines and Rca-T cell lines constructed by the method all come from single cloned tumor cells induced by SD pure mouse chemical carcinogens, and have the same genetic background and stable biological characteristics; and secondly, the characteristics of the cell lines such as hypodermal tumor formation in a nude mouse and high experimental liver transfer rate provide two practical animal models for study on prevention and treatment of oral mucosa epicytoma.

Described are compositions with tethered growth effector molecules, and methods of using these compositions for growing cells and tissues. Growth effector molecules, including growth factors and extracellular matrix molecules, are flexibly tethered to a solid substrate. The compositions can be used either in vitro or in vivo to grow cells and tissues. By tethering the growth factors, they will not diffuse away from the desired location. By making the attachment flexible, the growth effector molecules can more naturally bind to cell surface receptors. A significant feature of these compositions and methods is that they enhance the biological response to the growth factors. The method also offers other advantages over the traditional methods, in which growth factors are delivered in soluble form: (1) the growth factor is localized to a desired target cell population; (2) significantly less growth factor is needed to exert a biologic response. This method can be used as a means of enhancing the therapeutic use of growth factors in vivo and of creating surfaces which will enhance in vitro growth of difficult-to-grow cells such as liver cells.

The disclosure relates to a method for culture of a cis-platinum drug-resistant lung cancer organoid. The method comprises the following steps: mixing cis-platinum drug-resistant lung cancer tissue cells with the medium and matrix gel to obtain a to-be-cultured substance, wherein the medium contains matrix metalloproteinase-10, and the concentration of the matrix metalloproteinase-10 is 50-200nM based on the medium; and carrying out culture amplification on the to-be-cultured substance to obtain the cis-platinum drug-resistant lung cancer organoid. According to the culture method for the cis-platinum drug-resistant lung cancer organoid provided by the invention, since the employed medium contains the matrix metalloproteinase-10 with specific concentration, and the matrix metalloproteinase-10 with the specific concentration can promote in-vitro growth of the cis-platinum drug-resistant lung cancer organoid, the success rate of the culture is high when the method provided by the disclosure is used for the culture of the cis-platinum drug-resistant lung cancer organoid.

The invention provides a preparation method for chlorella antitumor polypeptide. The preparation method is characterized in that chlorella protein is extracted by using a low temperature ultrahigh pressure continuous flow cell crusher, then is hydrolyzed by trypsin and finally filtered with an ultrafiltration centrifuge tube so as to obtain chlorella polypeptide with a molecular weight in ranges of 0 to 3 KD, 3 to 5 KD, 5 to 10 KD and more than 10 KD. The chlorella polypeptide prepared in the invention has antitumor activity, e.g., the chlorella polypeptide has inhibitory effects on in vitro growth of human hepatoma carcinoma cells HepG2 and inhibition rates of chlorella polypeptide with a concentration of 1 mg / mL and in ranges of 0 to 3 KD, 3 to 5 KD and 5 to 10 KD on in vitro growth of hepatoma carcinoma cells (HepG-2) respectively reach 7%, 14% and 10%. Therefore, the chlorella antitumor polypeptide prepared in the invention is beneficial for development and utilization of antitumor health food and medicinal products.

Acceleration of botryococcenoids and growth by concomitant provision of appropriate light, minerals, and assimilable carbon. Specifically, methods, compositions and systems for the in vitro growth of hydrocarbons in photosynthetic organisms while maintaining a biologically exclusive monocultural environment, as for example, from Botryococcus species, is disclosed. Niche-nutrients can include about 200 ppm to about 3% nitrogen, and about 100 ppm to about 15% P205, and about 100 ppm to about 3.5% K20. In certain embodiments, the present invention relates to the growth of the Chlorophyta such as Botryococcus sp. in a nutrient medium that includes up to 15% phosphates, at least 3 ppm soluble iron, and up to about 70 ppm soluble zinc. Also disclosed is a substantially pure culture of Botryococcus braunii var. Showa, strain Ninsei, having the ATCC Accession No. PTA-7441, its parts, and hydrocarbons produced therefrom.

A method for supporting the in vitro growth of one or more eukaryotic cell type(s), the method comprising; seeding cells of said cell type(s) onto particles comprising basement membrane or a basement membrane-like substrate, and culturing said seeded cells in vitro under conditions suitable for expansion of said seeded cells, wherein said particles are less than about 500 μm in size.

The invention relates to a method for culturing a dedifferentiated or undifferentiated thyroid carcinoma organ. The method comprises the following steps: mixing dedifferentiated or undifferentiated thyroid carcinoma tissue cells with a thyroid carcinoma culture medium and matrix glue to obtain a to-be-cultured substance, wherein the thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin, and based on the thyroid carcinoma culture medium, the concentration of nicotinamide is 5-20 mM, and the concentration of BM-Cyclin is 5-30 [mu]g / ml; and carrying out culture amplification on theto-be-cultured substance to obtain the dedifferentiated or undifferentiated thyroid carcinoma organ. In the method for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, theadopted thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin with specific concentrations, so that the in-vitro growth of the dedifferentiated or undifferentiated thyroid carcinoma organ is promoted. Therefore, when the method provided by the invention is used for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, the culture success rate is relatively high.

The invention relates to a method for culturing liver cancer organoid. The method comprises the following steps: mixing liver cancer tissue cells with a culture medium and matrix gel to obtain a to-be-cultured substance, wherein the culture medium containing an insulin growth factor-2, and the concentration of the insulin growth factor 2 being 1-50ng / ml, preferably 5-10ng / ml based on the culture medium; and carrying out culture amplification on the to-be-cultured substance to obtain the liver cancer organoid. According to the culture method of the liver cancer organoid provided by the invention, the adopted culture medium contains the insulin growth factor-2 with the specific concentration, and the insulin growth factor-2 with the specific concentration can promotes in-vitro growth of theliver cancer organoid, so that the culture success rate is relatively high when the method provided by the invention is used for culturing the liver cancer organoid.

The invention discloses a medicament for curing breast cancer, which contains two components: arsenic trioxide and tetrandrine; the content of the tetrandrine is 5 to 40mg / ml; the content of the arsenic trioxide is 0.9 to 7.5mg / ml; the dosage of the tetrandrine is 5mg question mark kg<-1> question mark d<-> and the dosage of the arsenic trioxide is 0.3mg question mark kg<-1> question mark d<-1>; the components of the medicine also include fluorouracil and / or amylose. Another technical proposal of the invention is the medicament for inhibiting the in-vitro growth of breast cancer cells; and the content of the tetrandrine thereof is 0.5 to 4ug / ml and the content of the arsenic trioxide is 0.09 to 0.75ug / ml. The medicament for curing the breast cancer has the beneficial effects that the combined use of the arsenic trioxide and the tetrandrine can reduce the toxicity of the arsenic trioxide and the effect is better than that of independent medication; within the concentration range, the combined medication generates synergy which can be repeated; and if the concentration range is more or less than the concentration range, additive effect and even antagonism can be generated.

The invention relates to a group of micro RNA related to a BafilomycinA1 liver cancer and ovarian cancer inhibition cell line, belonging to the technical field of medical biology. The preparation method comprises the following steps: respectively acting BafilomycinA1 on a liver cancer cell line BEL-7402 and an ovarian cancer cell line HO-8910, and detecting the in-vitro growth inhibition, apoptosis promoting and invasion inhibition effects of BafilomycinA1 on two cells by performing soft agar cloning formation assay, electron microscopy observation, apoptosis related enzyme detection and in-vitro invasion assay; respectively acting 400nM BafilomycinA1 on the liver cancer cell line BEL-7402 and the ovarian cancer cell line HO-8910 for 48 hours, collecting dosing cells and control cells, extracting the RNA, analyzing the high-flux micro RNA between the drug group and the control group by using Affymetrix GeneChip Human Gene Array micro RNA array, and verifying the differences of the micro RNA by using qPCR, thereby obtaining co-sensitive micro RNA of the two cells. According to the group of micro RNA sensitive to the BafilomycinA1, on one hand, the micro RNA has the cancer inhibition effectiveness corresponding to the BafilomycinA1; on the other hand, the micro RNA has obvious change in two tumor cells, and a basis is provided for targets designed by taking the micro RNA as a potential broad spectrum anti-cancer drug.

Rapid purification of soluble protein of seashell nacre is carried out by: washing, acid treating, crushing, dialyzing, centrifuging, and drying. Furthermore, It is desalted with FPLC and is separated and purified by anionic exchange chromography. Within each cycle of 20min, 0.3-0.6mg purified soluble shell protein is obtained. It is used to induce human bone generation and to promote bone renovation in clinic. Meanwhile, it provides a technology of in vitro growth of valuable pearl.

Belonging to the fields of pharmacy and clinical pharmacy, the invention relates to a folic acid modified and vincristine encapsulated active targeting liposome drug delivery system, a preparation method and application in multidrug resistant tumor targeted drug delivery. The invention discloses a preparation method of a folic acid modified and vincristine encapsulated liposome. Cell specific uptaking and living animal imaging tests show that the drug delivery system has good in vitro and in vivo tumor cell targeting. 3-(4, 5-dimethylthiazole)-3, 5-diphenyltetrazolium bromide (MTT) analysis indicates that the drug delivery system has a good effect for inhibiting the in vitro growth of tumor cells. Pharmacodynamic results show that the drug delivery system has a good effect for inhibiting the in vivo growth of multidrug resistant tumors. The drug delivery system can enter the whole body blood circulation through intravenous injection dosing, then can be targeted to multidrug resistant tumor sites through tumor EPR effect and folic acid mediation and enter the tumor cells.

The invention discloses a culture medium capable of maintaining in-vitro growth of hyphae of ustilago esculenta, and belongs to the technical field of culture media. Every liter of the culture medium comprises the following components: 0.85 to 1.15 grams of K2HPO4, 0.45 to 0.55 gram of MgSO4.7H2O, 0.008 to 0.012 gram of FeSO4.7H2O, 0.45 to 0.55 gram of KCl, 2 to 3 grams of yeast powder, 4 to 6 grams of peptones, 3 to 4 grams of sucrose, 4.5 to 5.9 grams of histidine and 10 to 15 grams of agar. The culture medium capable of maintaining maintaining in-vitro growth of the hyphae of the ustilago esculenta is reasonable in compatibility; optimal carbon and nitrogen sources are added into a basic culture medium, so that in-vitro growth of the hyphae of the ustilago esculenta can be maintained by the culture medium.

The invention provides a disinfection composition for improving pig immunity and antiviral capability and a use method thereof, and the disinfection composition comprises a citrus extract and a nano zinc substance in a weight ratio of 1:0.5-10. Disinfectant prepared from the disinfection composition is combined with disinfection drinking water to disinfect individuals in vitro (growth environmentssuch as breeding houses, feeding troughs, excrement pools, individual appearances and the like) and in vivo (oral cavities, intestines, stomachs, blood and the like of pigs). By means of the internaland external firm mode, the negative effect problem caused by use of a common disinfectant can be solved, damage to skin, eyes and the like of individuals cannot be caused, comprehensive prevention and control over African swine fever are achieved, and spreading of diseases among the individuals is restrained. The disinfection composition has no influence on the respiratory system of an individual, has no damage to the skin of the individual, can purify the environment, improves the immune function of the organism, and can prevent and control African swine fever.