51 results about "Tissue/cell culture" patented technology

Disclosed is a scaffold including a semi-permeable membrane on an outer surface thereof. The present invention also discloses a method of preparing a scaffold covered with a semi-permeable membrane, including loading one or more scaffolds into a mold with a predetermined form and size; and adding a semi-permeable agent and a cross-linking agent to the mold and cross-linking the semi-permeable agent to form the semi-permeable membrane on the outer surface of each of the scaffolds. The scaffold covered with the semi-permeable membrane selectively introduces nutrients into the scaffold by allowing penetration of only external nutrients into the scaffold and excreting metabolic wastes generated by tissue cells to the outside of the scaffold. In addition, the scaffold has the morphology of a biological tissue of interest by cross-linking the small-sized scaffolds, thereby allowing uniform proliferation of tissue cells throughout the whole scaffold.

The invention provides a specific fluorescence probe substrates of human carboxylesterase 2 (CES2) and application thereof. The specific probe substrate is a benzoateb compound of a C4 hydroxyl naphthalimide, and is applicable to determine the enzyme activity of CES2 in a biological system. The CES2 enzyme activity determination flow comprises: selecting a hydrolysis benzoyl-removal reaction of the benzoate compound of the C4 hydroxyl naphthalimide as a probe reaction, and quantitatively determining the generation amount of a hydrolysis metabolite of the compound in a unit time, so as to determine the enzyme activity of CES2 in all biological samples, cells, bodies and integral organs. The probe is applicable to quantitative assessment of CES2 enzyme activity in biological samples of different species and different individual sources, and quantitative determination on CES2 enzyme activity in different sources of animal tissue cell culture fluids and cell preparation substances, so that the probe is expected to help to realize assessment on medicine disposal capability of important drug metablic enzyme CES2. Additionally, the probe also is applicable as an inhibitor for rapidly screening CES2 in vitro by means of the probe reaction.

The invention discloses a ratio type fluorescent probe substrate for cytochrome oxidase CYP1A and an application thereof. The specific probe substrate has a hydroxynaphthalimide alkane acid structure and can be used for determining the CYP1A enzymatic activity in a biosystem. A flow for determining the CYP1A enzymatic activity comprises the following steps: selecting a hydroxynaphthalimide alkane acid demethylation reaction as a probe reaction, and determining the CYP1A enzymatic activity in various biological samples by quantitatively detecting the production amount of demethylation metabolites in a unit time. The ratio type fluorescent probe substrate disclosed by the invention can be used for quantitative evaluation for the CYP1A enzymatic activity in biological samples of different species and different individual sources and for quantitative determination for the CYP1A enzymatic activity in different-source animal tissue cell culture fluids and cell prepared products, so as to realize evaluation for the medicine disposition capacity of the important medicine metabolizing enzyme CYP1A. In addition, the probe reaction can also be used for rapidly screening a CYP1A inhibitor in vitro and evaluating the inhibition capacity of the inhibitor.

The invention provides a specific probe substrate of catechol-O-methyltransgerase and application thereof. The specific probe substrate is a 7,8-dihydroxycoumarin compound and can be used for determination of activity of COMT enzyme in mammalian tissue and cells from different sources. The determination comprises the following concrete steps: selecting a coumarin compound having hydroxyl groups at positions 7 and 8 as a highly specific probe substrate; carrying out a COMT catalyzed reaction of the specific probe substrate in virtue of a COMT in-vitro incubation system; and determining the activity of COMT enzyme in each biological sample and each cell through quantitative detection of a product generation amount per unit time. The specific probe substrate can be used for quantitative evaluation of the activity of COMT enzyme in biological samples of different species and from different individual sources and quantitative determination of the activity of COMT enzyme in different-source-derived animal tissue cell culture fluids and prepared cell products, so assessment of capability of the important drug metablic enzyme COMT in disposition of drugs can be realized.

The invention provides specificity probe zymolyte of glucuronic acid transferase UGT1A1 and application. The specificity probe zymolyte includes dihydro flavonoids compounds of C-4' hydroxyl. Procedures of enzymatic determination are that C-4' hydroxyl-glucose aldehyde acid of bavachinin is selected to be metabolized to a probe reaction. Activity of UGT1A1 enzyme of each biological sample, cell, in vivo and whole organ is measured by quantitative determining of removing quantity of fructus psoraleae flavanone methyl ether in unit time or generation quantity of glucose aldehyde acidize metabolite of the fructus psoraleae flavanone methyl ether. The specificity probe zymolyte of glucuronic acid transferase UGT1A1 can be used for quantitative evaluating of the activity of the UGT1A1 enzyme of different species, different individual source biology samples and quantitative evaluating of the activity of the UGT1A1 enzyme of animal tissue cell culture fluid of different sources and cell prepared products so as to assess ability of handling drugs on important drug metabolic enzyme UGT1A1.

The invention provides a specific probe substrate for a cytochrome P450 3A4 enzyme and an application of the substrate in CYP3A4 enzyme activity measurement. The specific operation flow of the enzyme activity measurement comprises the following steps: carrying out the CYP catalytic reaction of the specific substrate by virtue of a CYP in-vitro incubation system by selecting any monomer in toad steroid series of compounds as a high-specificity probe substrate; and measuring the activity of the CYP3A4 enzyme in each biological sample and each cell by quantitatively detecting a product generated quantity or a substrate eliminated quantity in a unit time. The specific probe substrate provided by the invention can be used for quantitative evaluation of CYP3A4 enzyme activity in biological samples belonging to different species and deriving from different individual sources, and quantitative measurement of CYP3A4 enzyme activity in animal tissue cell culture fluids and cell products deriving from different sources.

The invention relates to a glycyrrhiza glabra callus cell culture method being capable of improving content of licoflavone. The method is characterized by including the steps of: (1) choosing full glycyrrhiza glabra seeds, and culturing the seeds on a MS basic culture medium containing 30 g / L of saccharose and 7 g / L of agar to obtain sterile seedlings, and inoculating the sterile seedlings on a callus induction culture medium for cultivation, and moving the sterile seedlings on a subculture medium for subculture; (2) selecting slight-yellow callus after the subculture in the step (1), and transferring the callus on a liquid culture medium to perform suspended shaking cultivation; (3) performing inductive cultivation during the suspended shaking cultivation process of the callus, and increasing culturing temperature from 25 DEG C to 35-45 DEG C from the sixth to twelfth day of the subculture period, and continuously culturing the callus for 1-3 days, reducing the culturing temperature to 25 DEG C, wherein coronatine is added during the cultivation at 35-45 DEG C and p-hydroxyphenylpyruvic acid is added when the temperature is reduced to 25 DEG C, and after the suspended shaking cultivation is finished, the glycyrrhiza glabra callus cell having high content of licoflavone can be obtained. The method achieves high content of the licoflavone.

The invention discloses application of a two-photon fluorescence probe for detecting cytochrome oxidase CYP3A4, and belongs to the technical field of biomedicine. The specific probe substrate can be used for measuring the enzyme activity of the CYP3A4 in a biological system. The procedure for determining CYP3A4 enzyme activity is as follows: Naphthalimide 4-position hydroxylation is selected as aprobe reaction, and the CYP3A4 enzyme activity in various biological samples can be determined by quantitatively detecting the amount of hydroxylated metabolites generated per unit time. The two-photon fluorescence probe can be used for quantitative evaluation of the CYP3A4 enzyme activity in the biological samples of different species and different individual sources, and quantitative determination of CYP3A4 activity in animal tissue cell culture fluids and cell preparations of different sources so as to realize evaluation of drug disposal capability of the important drug metabolizing enzymeCYP3A4. The two-photon fluorescence probe can also be used to rapidly screen inhibitors of the CYP3A4 in vitro, evaluate inhibitory ability of the inhibitors and detect the CYP3A4 activity in tumors,can detect the CYP3A4 activity in zebrafish, and can be used to detect drug-drug interaction of the CYP3A4 in vivo.

The invention belongs to a biological culture device, and in particular, relates to a tissue cell culture device comprising a tray; the tray is provided with fixing units for fixing culture dishes; the tray is movably arranged on a base; the base is provided with a circumferential limit unit for preventing the tray from circumferentially rotating; the bottom of the tray is horizontally provided with a rotary disk; the rotary disk is driven by a motor; the disk surface of the rotary disk is provided with two convex blocks which have smooth surfaces, wherein one of the two convex blocks is high and the other of the two convex blocks is low; the top parts of the two convex blocks make contact with the bottom surface of the tray and form sliding fit. In the rotation process, the upper convex block of the rotary disk can make the culture dishes on the tray periodically fluctuate and flip, a culture liquid in the culture dishes flows back and forth, and tissue cells at the bottoms of the culture dishes are intermittently exposed in air to meet the growth demand that the tissue cells need not only oxygen gas but also the culture liquid.

The invention provides a specificity probe substrate of glucuronic acid transferase UGT1A3 and application of the specificity probe substrate. Particular operation flows of enzyme activity measurement comprises the following steps of: selecting a bufanolide compound having 16 bits of hydroxyl to serve as the high specificity probe substrate, a UGT external incubation system is used for developing a UGT catalytic reaction of the specificity substrate, and UGT1A3 enzyme activity in each biological sample and cells is measured through product production quantity or substrate removing quantity in quantitative detection unit time. The specificity probe substrate can be used for quantitative evaluation of the UGT1A3 enzyme activity in the biological samples with different species and different individual resources and quantitative determination of the UGT1A3 enzyme activity in animal tissue cell culture fluid and cell preparation from different sources so as to achieve evaluation of capability of important drug metabolic enzyme UGT1A3 for handling drugs.

The invention discloses a culture medium for 3D culture of ovarian cancer tissue, the tissue is prepared from cytokine B27, N-acetylcysteine, R-spondin 1, A83-01, epidermal growth factors, glutamine,N2Supplement, estrogen, progesterone and vitamin E. The culture medium includes various cytokines and signal pathway regulatory factors which are needed by ovarian cancer tissue cell culture, mutually, directly and closely affect each other and coordinate with each other, so that the ovarian cancer tissue cells can better display the inherent active characteristics in the process of culture, and the overall characteristic highly similar to the living ovarian cancer tissue is achieved, and ovarian tumor cells subjected to 3D culture with the culture medium are clustered and are hypoxic in the middle and similar to the ovarian tumor tissue.

The invention provides a fluorescent probe substrate for testing of activity dipeptidyl peptidase IV and application of the fluorescent probe substrate and belongs to the technical field of biomedicine. The fluorescent probe substrate is a C-4 amide derivative GPAN of naphthalimide and can be used for testing enzyme activity of DPP-IV in different biological systems. A process for testing enzyme activity of DPP-IV includes: selecting GPAN amide hydrolysis reaction as probe reaction; quantitatively detecting generation quantity of a GPAN dipeptidyl-removed metabolism product to test activity of DPP-IV in various biological samples. The fluorescent probe substrate can be used for quantitatively testing enzyme activity of DPP-IV in biological samples different in species and individual source and enzyme activity of DPP-IV in animal tissue cell culture liquid and cell preparation products different in source, and is expected to realize quantitative evaluation on activity of DPP-IV which is a metabolic enzyme important to human body. In addition, with the help of probe reaction, the fluorescent probe substrate can be used for quickly screening inhibitors or inducers of DPP-IV and evaluating inhibiting or inducing capability thereof.

The invention discloses a face cream composition with a spot-fading effect and a preparation method thereof. The face cream composition is prepared from the following components in percentage by weight: 0.3-35.0% of a spot-fading functional component, 5.0-40.0% of an emollient, 0.5-30.0% of an emulsifying agent, 0.5-20.0% of a filling agent, 0.1-10.0% of a thickening agent, 0.3-5.0% of a preservative, 0.05-0.5% of EDTA-2NA, 0.01-1.0% of daily essence and the balance deionized water. A face spot-fading functional component is prepared by mixing an apple fruit / eryngium maritimum / rosa multiflora callus cell culture extract, a ceratium longissimum extract and sialic acid, wherein the apple fruit / eryngium maritimum / rosa multiflora callus cell culture extract is prepared by using a cell culture technology. The face spot-fading functional component can effectively inhibit generation of melanin, resist pigmentation and oxidation, balance the skin from inside to outside, fade face spots and make the skin fresh and bright.

The present invention relates to a method useful for topical compositions and the compositions. The topical compositions comprise centella asiatica extract, magnolia grandiflora bark extract or magnolia officinalis bark extract, hespyridine methyl chalcone, pea extract, citrus grandis peel extract, magnolia biondii flower extract, dipeptide-2, palmitoyl tetrapeptide-7, tripeptide-1, and optionally centella asiatica meristematic cell culture. The invention also provides methods thereof for application and production.

The invention discloses a method for identifying an anti-disease gene of an apple by combining apple callus tissue cell culture and genetic transformation, particularly provides a method for identifying a target gene as a disease resistance-related gene of the apple. An apple callus tissue is taken to serve as a transformation receptor, and the target gene is identified as the disease resistance-related gene of the apple. Proved by tests, apple callus tissue transformation is utilized for functional identification of the anti-disease gene of the apple, technical barriers, including long plant transformation period, difficulties in transformation and the like of the apple, in anti-disease gene function research of the apple are overcome, screening efficiency of the anti-disease gene of the apple is increased greatly, an efficient and simple method for screening and identifying the anti-disease gene of the apple is provided, and exploration of the anti-disease gene of the apple is enabled to be possible. The method has the advantages of efficiency, easiness in implementation and short consumed time, and establishes a base and a possibility for anti-disease gene screening and identification of the apple and in-depth research of a mechanism of action of the anti-disease gene of the apple.

The invention relates to a porous support for cartilage tissue engineering and a preparation method thereof, and belongs to the technical field of biological medical materials. The porous support is a three-dimensional porous support comprising carragheenan and chitosan and has porosity of 80-96% and aperture sizes of 50-500 microns. The preparation method comprises the following steps of preparing a carragheenan aqueous solution from distilled water, preparing a chitosan solution from a dilute acid solution, respectively beating the solutions, then blending the solutions, carrying out stirring, adding sodium carbonate crystal powder into the mixture, carrying out beating again, carrying out water-bath treatment, pouring the treated mixture into a stainless steel mold, carrying out freeze drying, carrying out immersion in a dilute acid solution, carrying out immersion in distilled water and carrying out vacuum drying to obtain the porous support for cartilage tissue engineering. The preparation method utilizes the raw material having wide sources and safety. The porous support has a certain function, high hardness, high porosity and no residue and is suitable for tissue cell culture or cartilage regeneration. The preparation method has simple and easy processes and a wide industrial prospect.

The invention relates to a tissue culture method of pinctada fucata, and belongs to tissue cell culture technology of the pinctada fucata. The method mainly comprises the following steps: (1) tissue mass taking, during which, carrying out surface sterilization of the pinctada fucata and cutting an adductor muscle in a sterile operating desk to take out a target tissue; (2) sterilization and cleaning, during which, wiping the mucus on the surface of the tissue by paper and transferring the tissue to tissue balanced salt solution after sterilization to clean surface mucus and residual antibiotics; and (3) tissue culture, during which, cutting the tissue mass into small pieces, putting the small pieces in a tissue culture dish and adding culture medium for culture. The tissue culture method has wide application range and better sterilization effect, and can protect tissue cell from being damaged; moreover, during tissue mass culture, the method ensures excellent adherence effect, vigorous cell migration, longer longevity in vitro and easy operation. The culture medium adopted by the method is more suitable for the culture of tissue cells of marine shellfish.

The present invention provides a culture device for tissue cell, and the culture device for tissue cell includes a tissue cell culture body. The porous material of the tissue cell culture body is composed by pore cavities which are classified according to the material pore size and cavity walls which are formed around the pore cavity at all levels. And the lower level small pore cavities are provided on the cavity wall formed around the upper level large pore cavities. The pore cavities within the same level are in communication with each other. And the pore cavities at all levels are also be in communication with each other. This device is particularly benefit for cell culture, so that tissue cells can freely and normally grow in three-dimensional space.

The invention discloses a specific fluorescent probe for glucuronyl transferase UGT1A1 and an application thereof. The substrate of the specific probe is (E)-2-(4-(4-hydroxystyryl)-3- cyano-5,5-dimethyl furan-2(5H)-ylene) malononitrile, which is named TCF for short and can be used for determining the enzymatic activity of UGT1A1 in a biosystem. A flow for determining the enzymatic activity of UGT1A1 is as follows: selecting a glucuronyl transferase acidification reaction of TCF acids as a probe reaction, and determining the activity of the UGT1A1 enzyme in various biological samples, cells, carriers and overall organs by quantitatively detecting the generation amount of glucuronyl transferase acidification metabolites in a unit time. The specific fluorescent probe disclosed by the invention can be used for quantitative evaluation for the activity of the UGT1A1 enzyme in different species and different individual sources of biological samples and for quantitative determination for the activity of the UGT1A1 enzyme in animal tissue cell culture fluids and cell prepared products from different sources, so as to realize evaluation for the medicine disposition capacity of the important medicine metabolizing enzyme UGT1A1. In addition, the probe reaction can also be used for rapidly screening a UGT1A1 inhibitor in vitro and evaluating the inhibition capacity of the inhibitor.

The invention discloses a multifunctional smart cell culture dish for cell culture, and relates to the technical field of culture dish structures. By means of design of three culture dish assemblies and installation of a temperature structure, the purpose of expanding teaching can be achieved. The multifunctional smart cell culture dish for cell culture comprises a functional rack. A base is of acircular cavity structure, and a column is vertically installed in the middle of the top surface of the base. Three culture dish bodies are uniformly arranged on the top of the base in an annular array manner, and a smart electrical box is also installed between two of the culture dish bodies. A temperature controller is in electrical control connection with three heating rings in a bottom cavityof the base by electrical connection wires. After tissue is cultured in the three culture dish bodies, states of the tissue are observed under a microscope, are compared mutually, and are also compared with changes formed during culturing. Therefore, when the multifunctional smart cell culture dish is applied to tissue cell culture in laboratory teaching, the purpose of flexible and expanded teaching can be achieved.

The invention relates to a method for inducing in-vitro transformation of scallop tissue cells, which comprises the following steps of constructing recombinant SV40LT and ientiviruses expression plasmids of a report gene GFP and packaging recombinant pantropic ientiviruses, which can express target genes, by using 293FT cells, and is characterized in that: the scallop tissue cells cultured in vitro, in particular chlamys nobilis heart cells cultured in vitro are infected with the recombinant pantropic ientiviruses, and transformation cells generating fluorescence or having splitting capability are obtained by blasticidin screening. In the invention, an exogenous gene is efficiently transferred to the chlamys nobilis heart cells cultured in vitro by the recombinant pantropic ientiviruses and stably integrated in a genome of the chlamys nobilis heart cells, so that the exogenous gene can be expressed continuously, cell proliferation and division can be promoted, and a basis for further constructing a chlamys nobilis cell system is laid.

Disclosed is a scaffold including a semi-permeable membrane on an outer surface thereof. The present invention also discloses a method of preparing a scaffold covered with a semi- permeable membrane, including loading one or more scaffolds into a mold with a predetermined form and size; and adding a semi-permeable agent and a cross-linking agent to the mold and cross-linking the semi-permeable agent to form the semi-permeable membrane on the outer surface of each of the scaffolds. The scaffold covered with the semi-permeable membrane selectively introduces nutrients into the scaffold by allowing penetration of only external nutrients into the scaffold and excreting metabolic wastes generated by tissue cells to the outside of the scaffold. In addition, the scaffold has the morphology of a biological tissue of interest by cross-linking the small-sized scaffolds, thereby allowing uniform proliferation of tissue cells throughout the whole scaffold.

The present invention relates to a tissue cell culture device which includes a tissue cell culture body. The porous material used as the tissue cell culture body is a porous metallic material which is formed by pore cavities classified by a pore size of the material and cavity walls surrounding to form the classified pore cavities. The cavity wall surrounding to form an upper level of large pore cavity is provided with a lower level of small pore cavity. The pore cavities in the same level are in communication with each other. The pore cavities in different levels are also in communication with each other. This device is particularly beneficial for cell cultivation and allows tissue cells to grow freely and normally in a three-dimensional space.

The invention discloses an NTDs (neural tube defects) rat embryo animal model and a constructing method thereof. Normal pregnant rats which have been pregnant for 6 to 10 days are injected with 19-23 mg / kg of retinoic acid at one time for intragastric administration intervention, then construction of the stable and efficient NTDs rat embryo animal mode can be induced rapidly after pregnant 10.5 days, and further, rat embryo brain vesicle tissue is separated out for cell culture while an NTDs cell model can be built. The animal model and the cell model constructed by the method can provide good platforms for research on NTDs molecular mechanism and cellular mechanism in vivo and vitro.

The invention relates to a separation method for mantle epithelial cells in vitro of pinctada martensii, belonging to the technical field of separation for histiocyte culture. The method mainly comprises steps of: (1) preparing Percoll gradient solution as follows: mixing nine parts of Percoll stock solution with one part of D-MBSS to obtain 100% of Percoll solution; and mixing the 100% Percoll stock solution with D-MBSS solution in different proportion to obtain the Percoll gradient solution; (2) preparing gradient layer as follows: previously using BSA solution to soak the tube wall of a centrifuge tube and preparing a gradient tube; (3) adding cell suspension as follows: adding cell suspension to be separated into the gradient tube; and (4) cell separation: after centrifugating, taking out cells, washing and heavy suspending. The invention can separate out single type of cell group from cells emigrated from primarily cultured tissue blocks, has low damage to cells, and can maintain activity of cells to the maximum extent. The cells after being separated still can be cultured. The operation is simple.