36 results about "Cell culture model" patented technology

The invention discloses hydrogel for three-dimensional culturing of tumor cells and applications thereof, and particularly relates to applications in fundamental research related to oncology and anti-tumor drug screening. The hydrolysis product of polymethyl vinyl ether maleic anhydride, i.e., methyl vinyl ether maleic anhydride copolymer, and a hydrogel cross-linking agent polyethylene glycol are used for a cross linking reaction to prepare the hydrogel suitable for being used as a three-dimensional culture medium of various tumor cells. Compared with commercialized BME gel and Collagenl gel available on the market, the hydrogel is determined to have the capabilities of simulating in-vitro environment and supporting three-dimensional growth of growth of cells in the culture medium through researching diversity factors of cell adhesion abilities and cell growth curves on tumor cell transfer and invasion, drug resistance experiments of anti-tumor drugs and the like, thus providing a three-dimensional cell culture model for researches on tumor molecular biology characteristics, drug screening and the like.

Endothelial implants restore vascular homeostasis after injury without reconstituting vascular architecture. Endothelial cells line the vascular epithelium and underlying vasa vasorum precluding distinction between cellular controls. Unlike blood vessels, the airway epithelium is highly differentiated and distinct from endothelial cells that line the bronchial vasa allowing investigation of the differential control tissue engineered cells may provide in airways and blood vessels. Through airway injury and cell culture models, tissue engineered implants of the bronchial epithelium and endothelium were found to promote synergistic repair of the airway through biochemical regulation of the airway microenvironment. While epithelial cells modulate local tissue composition and reaction, endothelial cells preserve the epithelium; together their relative impact was enhanced suggesting both cell types act synergistically for airway repair.

The present invention provides a cell-culturing model and a method for screening compounds which can be applied in treating or preventing hepatic cirrhosis. The cell culturing model comprises a population of hepatocytes and hepatic stellate cells (HSCs) derived from co-culturing, and at least one population of the cells comprises a nucleotide sequence fragment of a reporter gene and a cell specific regulatory sequence. The cell-culturing model of the present invention can be applied in high throughput screening for effective compounds of medication, and also in understanding the functional mechanism of the effective compounds.

The invention provides a method for establishing an in-vitro blood-brain barrier model based on a micro-fluidic chip. The micro-fluidic chip mainly comprises a cell inlet, collagen inlets and waste liquid outlets, wherein the cell inlet is communicated with the waste liquid outlets via two-dimensional channels, the collagen inlets are communicated with three-dimensional channels, and two-dimensional and three-dimensional microenvironment interfaces are formed at joints of the two-dimensional channels and the three-dimensional channels. The blood-brain barrier model is established on the base of the micro-fluidic chip. The method comprises the following steps: injection of collagen into the chip; and injection and culture of cells on the chip. Compared with conventional non-contact cell co-culture models or two-dimensional cell culture models, the invention has the following advantages: the method overcomes the problems of secondary inoculation and great time consumption of cell co-culture, solves problems in coexistence of two-dimensional and three-dimensional microenvironments needed by the physiological environment of blood-brain barriers, enables a microenvironment to be closer to the actual physiological microenvironment in the human body and can substantially reduce actual consumption.

The invention relates to a separation and culture method for different cellular components of a human mammary tissue, and belongs to the technical field of cell culture. The method comprises the following steps: (1) sampling a fresh human mammary tissue specimen; (2) carrying out mechanical shearing and collagenase and hyaluronidase combined digestion to prepare a mammary tissue cell suspension; (3) carrying out low-speed and differential centrifugation to layer and separate the different cellular components; (4) carrying out centrifugation, washing and adherent culture for multiple times to purify the cell components, so as to obtain the different cellular components of the mammary tissue, which comprise epithelial cells, matrix cells and preadipocytes. The separation and culture method disclosed by the invention can be used for quickly, simply and effectively separating and purifying the different cellular components from the same mammary tissue; the cultured mammary gland cells are sufficient in quantity, good in cell viability and purity which is up to 95% above; the separation and culture method can provide very useful materials for molecular biology study associated with mammary glands and breast cancer, and establishes a foundation for construction of a mammary gland microenvironment multi-cell culture model.

The invention discloses a layered co-culture method for intramuscular fat cells and skeletal muscle cells of a mammal. The layered co-culture method comprises the steps: after washing and cutting a longissimus tissue on the back of a butchered animal into pieces, digesting the longissimus tissue by using type-I collagenase, and sieving a digested mixed solution; collecting filtrate, centrifuging, respectively collecting mature fat cell sap on the upper layer and muscle cells on the lower layer, respectively adding a proper amount of serum-free medium to dilute and wash, and centrifuging; adding the centrifugate on the upper layer and the muscle cell sap on the lower layer into a culture flask with the capacity of 25cm<2>, and culturing in a culture box with the temperature of 37 DEG C and the CO2 content of 5%. The mature fat cells can float and adhere to a ceiling surface on the upper layer because of small buoyancy, while muscle cells sink to the bottom of the culture flask to grow. The cell culture model can be used for layering and co-culturing the intramuscular fat cells and the muscle cells of the same muscle tissue of an animal in the same culture solution, so that a novel in-vitro cell research model is provided for researching the influence of mature fat cell secreta to muscle cell differentiation or the interaction between the muscle cells and the fat cells.

A method for increasing or monitoring apoptosis in tumor cells by the co-administration of ionizing radiation and an anti-integrin antibody. Increasing apoptosis reduces tumor growth in vivo and in a cell culture model. The antibody is directed against the beta-1 integrin subunit and is inhibitory of beta-1 integrin signaling. Other molecules having an inhibitory effect on beta-1 integrin, either in signaling or in binding to its cognate extracellular receptors may also be used. The present method is particularly of interest in treatment of tumor cells associated with breast cancer, wherein radiation is currently used alone. The present method further contemplates a monoclonal antibody suitable for human administration that may further comprise a radioisotope attached thereto.

A method for increasing or monitoring apoptosis in tumor cells by the co-administration of ionizing radiation and an anti-integrin antibody. Increasing apoptosis reduces tumor growth in vivo and in a cell culture model. The antibody is directed against the beta-1 integrin subunit and is inhibitory of beta-1 integrin signaling. Other molecules having an inhibitory effect on beta-1 integrin, either in signaling or in binding to its cognate extracellular receptors may also be used. The present method is particularly of interest in treatment of tumor cells associated with breast cancer, wherein radiation is currently used alone. The present method further contemplates a monoclonal antibody suitable for human administration that may further comprise a radioisotope attached thereto.

The invention relates to a colorless and transparent sericin hydrogel, which is prepared by utilizing HRP and H2O2 to catalyze silk sericin cross-linking. The invention also provides application of the silk sericin hydrogel in biomedical materials. The invention also provides a stent obtained by freeze-drying of the silk sericin hydrogel. The invention also provides the role of the stent in in-vitro construction of a three-dimensional tumor model, in-vitro construction of a cell culture model, study of drug resistance of tumors, and screening of new drugs. The silk sericin hydrogel provided by the invention and the stent prepared therefrom can be used for carrying cells, especially tumor cells, and establishing a three-dimensional tumor model, and because of the colorless and transparent characteristics, the sericin hydrogel and the stent provided by the invention are conducive to observation and tracing of the constructed three-dimensional tumor model, thus better studying tumor cells and the development biological behaviors of tumor tissues.

The invention discloses a single-layer or multi-layer 3D brain glioma cell culture model, and a construction method and an application thereof. The construction method includes the steps: freezing a decellularized brain tissue, and cut into slices; freeze-drying the slices, and sterilizing to obtain a scaffold; and inoculating a single-slice or multi-slice slice scaffold with brain glioma cells to obtain a single-layer or multi-layer brain glioma cell 3D culture model. The used decellularized brain slice scaffold for constructing the 3D brain glioma model has the advantages of complete removal of cells and fewer nucleic acid residues, can partially preserve a stereoreticular structure and biological components of an original brain matrix, is suitable for growth of the brain glioma cells, and can simulate a microenvironment of the growth of the brain glioma cells in vivo; the multi-layer 3D brain glioma cell culture model can also simulate the hypoxia and nutrition gradient of glioma in vivo, is more close to the brain glioma cells growing in vivo, and is suitable for screening of antitumor drugs, co-culture of the brain glioma cells and tumor-related cells, and study on the mechanism of action between the brain glioma cells and ECM.

The present disclosure concerns an in-vitro full skin model which comprises a dermal equivalent and epidermal equivalent as well as from about 1 to about 100 three-dimensional sweat gland equivalents with respectively from about 500 to about 500000 sweat gland cells as well as a diameter of respectively from about 100 to about 6000 μm on a supporting layer. Furthermore, the present disclosure concerns the production of the full skin model as well as the use of this model as an in-vitro model, in screening methods as well as for in-vitro evaluation of the influence of cosmetic substances on the inhibition of sweat secretion as well as body odor.

A hepatitis C virus (HCV) genome is highly heterogeneous, and can be divided into eight genotypes and multiple gene subtypes according to the difference of genomic bases. HCV from patients cannot be cultured directly in cells in vitro. Infectious clones of HCV for constructing epidemic strains by a reverse genetic method is an important technical operation platform for promoting HCV basic researchand drug and vaccine research and development. The gene type 6 HCV is mainly prevalent in China and Southeast Asia. At present, no full-length infectious clones of the gene type 6 Chinese strain exist. The main content comprises establishment of in-vitro cell infectious clones CH6acc of the HCV gene type 6a in China and demonstration of application of the CH6acc in drug research. A CH6acc cell culture model can be used in HCV basic research, and drug and vaccine research and development.

The present invention provides a simple and robust human liver cell-based system in which persistent hepatitis C infection, persistent hepatitis B infection or ethanol exposure induces a clinical Prognostic Liver Signature (PLS) high-risk gene signature. The cellular model system for hepatocellular carcinoma (HCC) / cirrhosis development and progression may be used in the screening of compounds useful in the treatment and / or prevention of cirrhosis and / or HCC as well as in the identification biomarkers for the prediction of liver disease (especially cirrhosis) progression and HCC. The present invention also relates to specific compounds that have been identified, using such screening methods, as useful in the treatment and / or the prevention of HCC / cirrhosis.

The invention relates to a method for constructing a gene 2a type hepatitis c virus clinical isolates cell culture model. The invention further provides a recombined hepatitis c virus capable of efficient infection and reproduction in virus packaging cells. The technical scheme provided by the invention can be used for providing individual-based treatment for HCV patients, and developing vaccines with a wider spectrum of activity, filtrating specific anti-HCV drugs and the like.

The present invention relates to the field of in vitro 3D modeling of neural tissues, particularly of the brain. There is the need of developing cell culture models of neural tissue that reflect physiological aspects of neural tissue. The present invention provides methods of producing bioengineered neuronal organoids (BENOs) which form functional neuronal networks. The present invention also relates to uses and applications of the produced BENOs, e.g., in the fields of drug screening and personalized medicine.

The invention relates to a method for constructing a 3D model in vitro of colon cancer cell peritoneal metastasis. The method comprises the following steps: (1) uniformly mixing a fibroblast suspension with I type collagen, and adding the mixture to a culture vessel immediately; (2) placing the culture vessel at room temperature for 20 minutes, then placing the culture vessel in a cell culture incubator for 2 to 4 hours till the mixture coagulates, and spreading mesothelial cells into a plate for 18 to 38 hours; and (3) spreading colon cancer cells into the plate to complete the construction of the 3D model in vitro. The method can be used for simulating a physiological microenvironment for better understanding changes in adhesion and invasion ability during colorectal cancer metastasis, can improve an existing cell culture model in vitro for studying colorectal cancer metastasis and contributes to an overall understanding of the early metastasis of the colorectal cancer.

The present invention relates to the field of in vitro 3D modeling of neural tissues, particularly of the brain. There is the need of developing cell culture models of neural tissue that reflect physiological aspects of neural tissue. The present invention provides methods of producing bioengineered neuronal organoids (BENOs) which form functional neuronal networks. The present invention also relates to uses and applications of the produced BENOs, e.g., in the fields of drug screening and personalized medicine.

Endothelial implants restore vascular homeostasis after injury without reconstituting vascular architecture. Endothelial cells line the vascular epithelium and underlying vasa vasorum precluding distinction between cellular controls. Unlike blood vessels, the airway epithelium is highly differentiated and distinct from endothelial cells that line the bronchial vasa allowing investigation of the differential control tissue engineered cells may provide in airways and blood vessels. Through airway injury and cell culture models, tissue engineered implants of the bronchial epithelium and endothelium were found to promote synergistic repair of the airway through biochemical regulation of the airway microenvironment. While epithelial cells modulate local tissue composition and reaction, endothelial cells preserve the epithelium; together their relative impact was enhanced suggesting both cell types act synergistically for airway repair.

The present invention provides a cell-culturing model and a method for screening compounds which can be applied in treating or preventing hepatic cirrhosis. The cell culturing model comprises a population of hepatocytes and hepatic stellate cells (HSCs) derived from co-culturing, and at least one population of the cells comprises a nucleotide sequence fragment of a reporter gene and a cell specific regulatory sequence. The cell-culturing model of the present invention can be applied in high throughput screening for effective compounds of medication, and also in understanding the functional mechanism of the effective compounds.

The invention discloses a coccidia cell culture model. The coccidia cell culture model is built through the processes that sporozoites of Eimeria tenella are inoculated into DF-1 cells of a Chicken Embryo Fibroblast Cell line and grow into first generation merozoites. The invention also discloses an application of the coccidia cell culture model in a selection of anticoccidial drugs. The coccidia cell culture model can be utilized for establishment of a test method for measuring objectively sporozoite invasion activity, and is suitable for a selection of anticoccidial drugs and researches of gene or protein functions during a coccidia growth process.

The invention discloses a carotid bifurcation three-dimensional cell culture model for drug screening, and a construction method thereof. Human-derived cells are used to make up for the defects of different species of animal models, and are closer to the real microenvironment of human bodies than traditional two-dimensional planar environments. The method can reflect the conditional controllability and intuitiveness of the traditional 2D cell culture, and can link a 2D cell culture system with tissue and organ researches to effectively simulate the influences of different pathological stimulion in-vivo cell functions, so the evaluation of the cell functions is accurate; and compared with existing three-dimensional vascular structure culture models, the model constructed by the method cangive stimulating factors in different pathological environments of simulated human bodies in order to study the changes of the form and functions of the cells in a disease state.

The invention relates to use of galangin and a derivative thereof in preparing a medicine or a feed for resisting coccidiosis. Compared with the prior art, the galangin and the derivative thereof have an obvious inhibiting effect on eimeria tenella. The inhibiting effect is reflected in that the galangin and the derivative thereof can effectively inhibit reproduction of eimeria tenella on a cell culture model of the eimeria tenella. Besides, the inhibiting effect is verified by an animal body test. Aiming at the discovery of the galangin and the derivative thereof, use fields of the galangin and the derivative thereof are widened, and a new scheme is provided for development of anticoccidial drugs.

The invention relates to the technical field of animal disease prevention and control, in particular to a novel application of cynarin, and particularly relates to an application of cynarin in preparation of drugs or feeds for resisting coccidiosis. Compared with the prior art, the invention has the following beneficial effects that cynarin has an obvious inhibition effect on coccidium (especially eimeria tenella), and the inhibition effect is reflected in that the cynarin can effectively inhibit the reproduction of the eimeria tenella on an MDBK cell culture model of the eimeria tenella. Besides, the inhibition effect is verified by animal body tests. With the novel application of cynarin, the application field of cynarin is widened, and a new scheme is provided for development of anticoccidial drugs.

The invention discloses a cytoskeleton as well as a construction method and application thereof. The method comprises the following steps: mixing matrigel and mesenchymal cells to obtain cell gel, and culturing the cell gel to obtain the cell scaffold. The invention provides a simple, rapid and low-cost method for constructing the cell scaffold, the prepared cell scaffold has a latticed structure, the pore diameter is about 50-500 microns, the anatomical diameters of various glands and vessel tissues of a human body are covered, and the cell scaffold can be used as a growth framework of various tissue organ cells of the human body, such as kidney, prostate, micro / small blood vessels or testis, so that the cell scaffold has a good application prospect. The method is used for preparing the 3D cell culture model, and the constructed 3D cell culture model can better simulate an in-vivo physiological structure.

The present disclosure relates to an in-vitro method for identifying and analyzing ion channels and / or water channels and / or receptors of signal transduction, in which a three-dimensional sweat gland equivalent having from about 500 to about 500,000 sweat gland cells and a diameter of from about 100 to about 6,000 μm is firstly provided and then any ion channels and / or water channels and / or receptors of signal transduction present in this equivalent are infected and analysed. In a further method step c) the influence of test substances on the proteins identified previously in step b) is examined. Since the three-dimensional sweat gland equivalents used in step a) comprise differently differentiated cells and portray the in-vivo situation well, the measurement data obtained with the in-vitro method as contemplated herein can be transferred well to the in-vivo situation.

A method for the in-vitro investigation of sweating processes, in which a three-dimensional sweat gland equivalent is stimulated adrenergically, and the reaction of the three-dimensional sweat gland equivalent is determined by measuring the cyclic adenosine monophosphate (cAMP) content.

The present invention is extracorporeal cell culture model and process for HIV-1 infected CD4+ cell, and relates to life science and medicine field. The present invention provides comprehensive HIV-1 infection treating mechanism for curing HIV-1 infection radically, designs new HIV-1 antagonizing medicine, provides new detection mechanism for detecting HIV-1 infection treating medicine, and provides relevant experiment reference. The present invention may be used in screening new HIV-1 antagonizing medicine for curing AIDS in early stage radically.

The invention discloses a high-throughput automatic continuous cell culture method and application thereof. The method comprises the following steps: (1) conducting inoculation and culture on a provided cell culture; (2) conducting cell sedimentation, and removing supernate in reactors; (3) adding a fresh culture medium; (4) calculating a release rate (B) according to the density of cells in the supernate, and maintaining the density of the cells; and (5) repeating the steps (2) to (4) until continuous cell culture is completed. According to the high-throughput automatic continuous cell culture method and application thereof, compared with a traditional perfusion reactor, a provided continuous cell culture model is high in throughput, high in automation degree, small in culture volume andlow in cost; and compared with culture by means of shaking, the high-throughput automatic continuous cell culture method has strong rector characteristics, strong culture result representation, low probability of personal errors and high reliability. Moreover, an additional cell retention device in the perfusion reactor does not need to be used in the high-throughput automatic continuous cell culture method, so that the problem that the cell retention device is liable to be blocked is solved.

A research model for monitoring demyelination or remyelination in a sample of cells that comprises providing cells, typically CNS cells, and contacting cells with a demyelination solution such as one that includes one of hexachlorophene and / or lysophosphatidylcholine.

The invention relates to the technical field of animal disease prevention and control, and particularly relates to novel application of deoxyrhaponticin, and specifically relates to application of deoxyrhaponticin in preparation of anti-coccidiosis drugs or feed. Compared with the prior art, the invention has the following beneficial effects: deoxyrhaponticin is found to have an obvious inhibitory effect on coccidium, the inhibitory effect is reflected in that deoxyrhaponticin can effectively inhibit the reproduction of Eimeria on a cell culture model of Eimeria, and the inhibitory effect is verified by animal experiments. Aiming at the discovery of deoxyrhaponticin, the application field of deoxyrhaponticin is broadened, and a new scheme is provided for the development of anti-coccidiosis drugs.