261 results about "Matrigel" patented technology

The present disclosed subject matter relates to methods and compositions for restoring a diseased or damaged tooth such that infection is inhibited or eliminated and pulp regeneration is facilitated. The disclosed subject matter also includes a composition comprising a physiologically acceptable matrix seeded with pulp cells. The matrix can be capable of being injected into the pulp chamber of a tooth. In some embodiments, the matrix of a composition includes a hydrogel (e.g., collagen, chitosan, alginate, MATRIGEL™, gelatin, JELL-O®, fibrin), a mesh (e.g., polylactide-coglycolide (PLGA) mesh, polylactide (PLA) mesh, or polyglycolide (PGA) mesh, a cross-linked fiber mesh, a nanofiber mesh, a mesh fabric, biodegradable polymer mesh), a microsphere (biodegradable polymer microsphere, a hydrogel microsphere), or a combination of any of the foregoing. In yet other embodiments, the matrix includes a nanofiber, an artificial three-dimensional scaffold material, or a synthetic three-dimensional scaffold material.

The present invention is drawn to the generation of micropatterns of biomolecules and cells on standard laboratory materials through selective ablation of a physisorbed biomolecule with oxygen plasma. In certain embodiments, oxygen plasma is able to ablate selectively physisorbed layers of biomolecules (e.g., type-I collagen, fibronectin, laminin, and Matrigel) along complex non-linear paths which are difficult or impossible to pattern using alternative methods. In addition, certain embodiments of the present invention relate to the micropatterning of multiple cell types on curved surfaces, multiwell plates, and flat bottom flasks. The invention also features kits for use with the subject methods.

A three-dimensional microwell system that supports long term pluripotent cell culture and formation of homogeneous embryoid bodies (EBs) is described. Microwell-cultured pluripotent cells remain viable and undifferentiated for several weeks in culture and maintain undifferentiated replication when passaged to Matrigel®-coated, tissue culture-treated polystyrene dishes. Microwell-cultured pluripotent cells maintain pluripotency, differentiating to each of the three embryonic germ layers. Pluripotent cell aggregates released from microwells can be passaged for undifferentiated replication or differentiated to monodisperse EBs. The ability to constrain pluripotent cell growth in three dimensions advantageously provides for more efficient, reproducible culture of undifferentiated cells, high-throughput screening, and the ability to direct pluripotent cell differentiation by generating monodisperse EBs of a desired size and shape. Cardiomyocyte-rich EBs are obtained from pluripotent cells cultured in microwells of defined size and shape.

We disclose a particle comprising a matrix coated thereon and having a positive charge, the particle being of a size to allow aggregation of primate or human stem cells attached thereto. The particle may comprise a substantially elongate, cylindrical or rod shaped particle having a longest dimension of between 50 μm and 400 μm, such as about 200 μm. It may have a cross sectional dimension of between 20 μm and 30 μm. The particle may comprise a substantially compact or spherical shaped particle having a size of between about 20 μm and about 120 μm, for example about 65 μm. We also disclose a method of propagating primate or human stem cells, the method comprising: providing first and second primate or human stem cells attached to first and second respective particles, allowing the first primate or human stem cell to contact the second primate or human stem cell to form an aggregate of cells and culturing the aggregate to propagate the primate or human stem cells for at least one passage. A method of propagating human embryonic stem cells (hESCs) in long term suspension culture using microcarriers coated in Matrigel or hyaluronic acid is also disclosed. We also disclose a method for differentiating stem cells.

The invention relates to a culture medium for a normal epithelial cell of a human or mammal, primary separation culture, subculture, 3D gas-liquid culture and 3D matrigel culture methods, the normal epithelial cell generated by using the culture medium and the culture methods and application of the normal epithelial cell to a toxicological evaluation system. The culture medium is prepared by mixing DMEM and Ham's F-12NUTRIENT MIX according to the volume ratio of 3:1 and also adding 4-6% of fetal calf serum, 1-3nM triiodothyronine, 0.4-0.65% of insulin-transferrin-selenium reagent, 4-6mu g / ml transferrin, 9-11ng / mL epidermal growth factors, 0.3-0.5mu g / mL hydrocortisone, 0.5-1.5nM cholera toxin, 0.4-0.6mu g / mL amphoterrible B, 35-45mu g / mL gentamicin, 45-55nM calpeptin, 35-45ng / ml recombinant human IL-1RA and 3mu g / ml recombinant human R-Spondin-1. The culture medium disclosed by the invention can be used for carrying out separation culture or subculture on the normal epithelial cell of the human or the mammal and any other various tissue source, rapidly proliferating the normal epithelial cell in vitro and establishing a cell line; and the normal epithelial cell is a normal diploid cell and is applied to the toxicological evaluation system of the human or the mammal.

A three-dimensional microwell system that supports long term embryonic stem cell (ESCs) culture and formation of homogeneous embryoid bodies (EBs) is described. Microwell-cultured ESCs remain viable and undifferentiated for several weeks in culture and maintain undifferentiated replication when passaged to Matrigel®-coated, tissue culture-treated polystyrene dishes. Microwell-cultured ESCs maintain pluripotency, differentiating to each of the three embryonic germ layers. ESC aggregates released from microwells can be passaged for undifferentiated replication or differentiated to monodisperse EBs. The ability to constrain ESC growth in three dimensions advantageously provides for more efficient, reproducible culture of undifferentiated cells, high-throughput screening, and the ability to direct ESC differentiation by generating monodisperse EBs of a desired size and shape.

The invention discloses tissue-engineered neural tissues and a construction method thereof, belonging to the technical field of tissue engineering and regeneration medicine. Subcultured and purified neural stem cells serve as seed cells of the tissue-engineered neural tissues, a liquid type I collagen and Matrigel matrix serves as stent materials, and the seed cells and the stent materials are cultured after being compounded in vitro. A stent system adopted in the invention can be used for effectively maintaining the survival and proliferation of the neural stem cells, promoting the differentiation of the neural stem cells, and forming engineered neural tissues consisting of mature neurons, astrocytes and oligodendrocytes. The tissue-engineered neural tissues have an important significance in treating neural system injury through the transplantation of the tissue-engineered neural tissues, and can also serve as an in-vitro model for the psychological research of neurodevelopment, cerebral trauma and neuroelectricity.

The invention discloses an in-vitro recombined human skin epidermis model.A layer of hemidesmosome protein is arranged on the lower surface of a basal cell layer, the upper surface of the basal cell layer is covered with 6-8 spinous cell layers containing short protruding spinous cells, the upper surface of the spinous cell layers is covered with 3-4 granular cell layers containing transparent granular cells, the upper surface of the granular cell layers is covered with a keratinocyte layer, all the cell layers are obviously separated, and the overall barrier function of a lipid composition is approximate to that of human skin.A preparation method comprises the steps of preparation of matrigel particles, screening and amplification of epidermal stem cells, preparation of the basal cell layer, preparation of the spinous cell layers, preparation of the granular cell layers, and maturation and keratinization of the in-vitro recombined epidermis model.The model is applied to evaluation of cosmetics with the human skin barrier repair effect, screening of plant extract with the human skin barrier repair effect, evaluation and detection of irritation of medical apparatus materials on the human skin and detection of irritation of chemicals on the skin.

The invention provides a primary cell culture medium for culturing primary esophageal squamous carcinoma epithelial cells and containing a combination of an MST1 / 2 kinase inhibitor and a ROCK kinase inhibitor, and a culture method using the primary cell culture medium. According to the culture method, the primary cells are cultured on a culture vessel coated with extracellular matrigel by using the primary cell culture medium, so that the primary cells are rapidly proliferated. The cell model obtained by the primary cell culture medium and the primary cell culture method can be used for evaluating and screening curative effects of medicines.

The invention provides an in vitro culture method of cancer type organs. According to the embodiment of the invention, the method comprises the steps of performing shearing and digestion treatment oncancer tissue samples, wherein the size of cancer tissue after the shearing treatment is 1-2mm<3>, and the digestion treatment is performed under the action of combined digestive enzymes; and performing re-suspension treatment on cancer cells obtained after digestion treatment with precooling substrate glue, wherein the concentration of the cancer cells in the substrate glue is 500-1500 / 10 [mu]L;and performing point and pallet treatment on cancer cell suspension, and performing culture treatment on the cancer cells after point and pallet treatment is performed, so as to obtain cancer organs.According to the method of the embodiment of the invention, the enzymolysis efficiency of the cancer cells is improved, the success rate of subculturing of organoids is increased, and long-term culture of the organoids can be realized, and a biology sample database can be established.

Cell transplantation for constructing biological heart pacemaker is used embryo source dry / ancestral cell and adult mesogalia dry cell as cell sources. The process is carried out by oriented differentiation inducing for heart pacemaker cell by embryo source multifunctional dry cell, bone marrow adhering to obtain primary culture cell, clone culturing to obtain purifying system by diluting, applying paracrine factor endothelin-1 or nervous adjusting protein-1, extracellular matrix laminated adherent protein and fiber connecting protein etc. substrate glue, trans-dyeing pacemaker gene HCN4, and in-vitro inducing embryo source multifunctional dry cell. It is various and non-immunogenicity. It can be used to treat sick sinus syndrome.

The invention discloses a method for in-vitro separation and culture of goat male germ stem cells. Spermatogonia are obtained from the testicular convolutedtubules of a male goat. A method for separating and purifying the goat male germ stem cells adopts 0.2% of gelatin and a Matrigel differential adherence combined cloning method to differentiate and separate un-adhered goat male germ stem cellsfrom other cells for adherence culture; and the culture of the separated goat male germ stem cells is MEF feeder-layer culture of feeder-layer-free culture. The separated goat male germ stem cells have ES cell characteristics and the potential of differentiation into nerve cells, cardiac myocytes and sperm sample cells.

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 relates to a co-culture model of microencapsulated tumor cells and endothelial cells. The preparation method comprises the steps: first, tumor cells are mixed into 2 percent of sodium alginate solution to lead cells suspension which is injected with 100mmol / L of calcium chloride solution to be colloidized through a microcapsule generator then; normal saline is used for washing the obtained material which then reacts with 0.1 percent of polylysine solution and is washed by normal saline; next, 0.15 percent of sodium alginate solution is added and washed by normal saline; the 55mmol / L sodium citrate solution is employed to liquefy microcapsule core and cultured in a CO2 of culture box; the microencapsulated tumor cells are directly used for being cultured or frozen reserved; the endothelial cells are employed to be adherently cultured or led to three dimensional growth; the microencapsulated tumor cells are added into the same culture solution; co-culture is done; tumor cells and endothelial cells are separated; finally, travelling gene detection, protein detection and culture liquid cytokine detection are done. The invention overcomes the limitation of Matrigel method and the defect of Transwell method, has an imitated micro-environment for microencapsulated tumor cells and endothelial cells to interact, and has long service life for one-time making and decreases research personnel and financial resources.

The invention discloses a method for separation and in-vitro culture of pancreatic cancer tissue organs of human. The method comprises the following steps that (1) after the pancreatic cancer tissue of human is cut, enzymic digestion is repeatedly carried out for 10-20 minutes at 37 DEG C in a rotation speed of 30-40 rpm three times, and a pancreatic cancer single cell is obtained; (2) the pancreatic cancer single cell and matrigel are uniformly mixed, then a culture medium containing a growth factor PEG2 and gastrin I are added for culture, and the pancreatic cancer tissue organs of human areobtained. The method for separation and in-vitro culture of the pancreatic cancer tissue organs of human has the advantages that the tumor tissue is digested by adopting a short-time repeated digestion method, which can effectively improve the cell yield and cell viability; the culture medium containing the growth factor PEG2 and the gastrin I is adopted for culture, which can effectively promotethe growth of the pancreatic cancer organs and increase the survival rate of the pancreatic cancer organs.

The invention discloses an ovarian cancer tissue 3D cultivation method. The method includes the steps that matrigel is dripped into a pore plate, and standing is conducted at an indoor temperature or in a CO2 cultivation box so that the gel can be solidified; a tumor tissue block submerged in a cultivation solution is moved onto the solidified matrigel, then matrigel is dripped onto the tumor tissue block and the gel is solidified in the same way; a cultivation solution is added for cultivation for 1-3 days; the tumor tissue block is taken out and fixed. Chemotherapeutics can be added during cultivation, the tissue block is fixed after cultivation, the cell shape is observed after dyeing, and the nucleus malformation rate is calculated. By means of the established cultivation method and detection indexes, the pharmacological effects and the function characteristics of anti-tumor drugs can be evaluated more quickly and accurately, the technical defects of cell 2D cultivation and animal experimental evaluation can be overcome, and it is hopeful that the research and development of innovative anti-cancer drugs in China and the precision therapeutic process can be promoted.

The invention provides a high-flux cultural method for an organoid type spheroid. The high-flux cultural method comprises the following steps of (1) respectively injecting cell-containing matrigel andfluorocarbon oil into a tee device to obtain the organoid type spheroid; and (2) heating the organoid type spheroid obtained in the step (1), linking the heated organoid type spheroid with a 3D printing platform, distributing the organoid type spheroid, and then culturing the organoid type spheroid. According to the high-flux cultural method provided by the invention, through micro-fluidic, an organoid is obtained, through combination with the 3D printing platform, the organoid can be simply, quickly and accurately printed to a culture carrier, the size and the structure of the organoid are appropriate and uniform and controllable, the number of the organoids in a culture cavity of each culture carrier is unchanged, and besides, drug susceptibility test and toxicity test are realized. Clinical pretest is accurate, the application prospect is favorable, and the high-flux cultural method has high application value.

The invention provides a primary cell culture medium for culturing primary mammary epithelial cells and containing amphiregulin, and a culture method using the primary cell culture medium. Through the culture method, the primary cell culture medium is used for culturing primary cells on a culture vessel coated with extracellular matrigel, the primary cells grow on the culture vessel coated with the extracellular matrigel, and the primary cells are rapidly proliferated under the combined action of nutritional factors contained in the primary cell culture medium and extracellular matrigel. A cell model obtained by the primary cell culture medium and the primary cell culture method can be used for curative effect evaluating and screening of medicines.

The invention relates to an animal model establishment method of human primary skin squamous epithelial cell cancer stem cell. The method comprises the following steps of: preparing collected skin squamous epithelial cell cancer tissue in a clinical case into a single-cell suspension, separating CD31-CD24-CD45-CD61-CD133+tumor stem cell population, suspending and mixing with 1*10<6> human primary mouse embroy fibroblasts and 1*10<6> human primary endothelial cells in matrigel, and then injecting the obtained product into microenvironment through humanized nude mouse subcutaneous tumorigenesis parts. The invention makes further research on molecular biological mechanisms of human skin primary SCC tumour possible.

The invention discloses an in-vitro separating and culturing method for a germline stem cell. The method comprises the following steps of: obtaining a cell from germinal epithelium of an ovary of a female mammal, differentiating and separating a non-adherent germline stem cell from other adherent cultured cells by adopting gelatin with the concentration 0.2 percent and a Matrigel differential adherence combination cloning method; and culturing the germline stem cell obtained by separating on an MEF (Mouse Embryonic Fibroblast) feeder layer. The separated germline stem cell has ES (Embryonic Stem) characteristic and the potential of differentiating into a muscle cell, an adipose cell and oocyte-like cell and various cells with three internal germinal layers.

The invention discloses microencapsulated human pancreatic carcinoma cells, and a preparation method and application thereof. The preparation method comprises the following steps of: adding suspension with human pancreatic carcinoma cells into Matrigel-containing sodium alginate solution; spraying into a calcium chloride solution in high voltage electrostatic environment to obtain microspheres; and performing ion exchange reaction on the microspheres and a sodium citrate solution, and thus obtaining the microencapsulated human pancreatic carcinoma cells. The obtained microencapsulated human pancreatic carcinoma cells have uniform size and the diameter of about 420mm. Cells in microcapsules are well proliferated, and grow in a three-dimensional way. The microencapsulated human pancreatic carcinoma cells are applied to subcutaneous tumor and in-situ tumor animal models, and have the advantages of high modeling success rate and quick tumor growth compared with the traditional tumor cell suspension injection method.

The invention discloses a novel medium for three-dimensional tumor cell culture. The three-dimensional cell culture medium can be used for wrapping tumor cells for three-dimensional culture so that anti-tumor drugs can be further screened. According to preparation of the three-dimensional cell culture medium, a sodium alginate solution, a sodium hyaluronate solution and an extracellular culture medium Matrigel are mixed in a proper proportion, and then cells are wrapped. Sodium alginate can be used for directly wrapping cells, but is bad for proliferation and differentiation of tumor cells, while Matrigel is an ideal cell three-dimensional culture medium bu is high in price, poor in mechanical strength and bad for screening and pharmacological research of anti-tumor drugs. The mechanical property of the medium is improved; meanwhile, formation of the tissue structure of extracellular culture cells is effectively promoted, usage of Matrigel is reduced or replaced, the cost of tumor cell three-dimensional culture is greatly reduced, xenograft and animal experiments can be conveniently carried out on three-dimensional cells, and the preparation method is simple and high in practicability.

The present invention provides a novel composition for treating angiogenic diseases using an extracellular matrix membrane of a cartilage-derived cell having a membrane or film form, which is excreted from cartilage cells that are cultured when the cartilage cells isolated from the cartilage of an animal is monolayer-cultured, and a transplant material for the cornea or the conjunctiva. The composition for treating the angiogenic diseases and the transplant material for the cornea or the conjunctiva, according to the present invention, suppresses endothelial cells from attaching or proliferating and impedes the movement of endothelial cells, thereby suppressing vascular endothelial growth and vascular penetration and thus ultimately suppressing angiogenesis. [Reference numerals] (AA) Matrigel; (BB) Matrigel / CD-ECM powder; (CC) Matrigel / Amnion powder

The present invention belongs to the field of biomedicine, and relates to a nanometer fiber polymer composite membrane, a preparation method and uses thereof. According to the nanometer fiber polymer composite membrane, the low layer is a hydrogel nanometer fiber membrane, the middle layer is a Matrigel matrix gel layer, and the upper layer is a retinal pigment epithelium layer. The nanometer fiber polymer composite membrane can be adopted as a bio-scaffold for supporting growth and transplantation of RPE cells, and has characteristics of softness, elasticity, good permeability, biodegradability and a structure similar to extracellular matrix, wherein adhesion and differentiation of the RPE monolayer cells and surgical procedure operations are easily achieved with the characteristics, and the nanometer fiber polymer composite membrane can be discharged out of a body through a metabolism process after transplantation so as not to generate harm to the human body. According to the present invention, a bio-nanometer material and cell engineering are combined, brightness and hope are provided for age-related macular degeneration (ARMD) patients with the successful research, and great economic benefits and social benefits are generated.

The invention provides a method for culturing and subculturing intestinal cancer organoid derived from circulating tumor cells. The method comprises the following steps: (1) mixing a human colorectalcancer circulating tumor cell organoid culture medium with Matrigel according to a ratio of 1:1 to prepare a starting plate glue; (2) resuspending cells captured from the blood of a metastatic colorectal cancer patient by a cell filter with the pore diameter of 5.5 [mu]m by using the starting plate glue, and dropwise adding the cells into a 24-pore plate; (3) after the starting plate glue is solidified, adding the human colorectal cancer circulating tumor cell organoid culture medium around the starting plate glue, and culturing; (4) during subculturing, discarding the original culture medium,and adding a TrypLE digestive juice for digestion; and (5) stopping digestion by using a DMEM containing 10% of FBS, cleaning twice by using PBS, and re-suspending a starting plate by using the starting plate glue. The method for culturing and subculturing intestinal cancer organoid derived from the circulating tumor cells provides an experimental model for research on metastasis and drug resistance mechanisms caused by the circulating tumor cells, can perform drug sensitivity screening, and provides an effective basis for individualized treatment of advanced intestinal cancer.

The invention relates to the technical field of organoid culture, in particular to a culture medium for preserving organoids at room temperature and a method for maintaining the growth activity of the organoids. The culture medium for preserving the organoids at room temperature comprises a DMEM high-glucose culture medium, Glutamax, HEPES, antibiotics, Matrigel, additives, a TGF-beta inhibitor and a Wnt signal pathway activator, and the volume fraction of the Matrigel is 20%-25% in the culture medium for preserving the organoids at room temperature. The culture medium provided by the invention can improve the biological activity of organoid preservation, and the method for maintaining the growth activity of the organoids can ensure that the organoids can maintain the activity for the longest time at room temperature, the growth activity and morphology of the organoids are not influenced, the activity can be quickly recovered after the organoids are transferred into a cell incubator, the growth of the organoids is continued, and the dissociation phenomenon is avoided.

The present invention relates to matrices (e.g., fibrin-alginate matrices; fibrin-alginate-matrigel matrices) for culture of cells, organs (e.g., ovary or fragment thereof), cells and cell aggregates (e.g., ovarian follicles, embryoid bodies), and tissues. In some embodiments, protease inhibitors e.g., aprotinin) are used to prevent the degradation of fibrin.

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 discloses a mmethod for obtaining analogous retinal tissues rich in cone and rod cells by using human induced pluripotent stem cells. The method comprises the following steps: digestinghiPSCs to obtain cell pellets, and then carrying out suspension cultivation on the cell pellets to obtain an embryoid body; reinoculating the embryoid body into a culture dish enveloped by Matrigel, and carrying out induced differentiation inside an induced cultivation solution to obtain nerve retina (NR) and RPE; stirring up the NR and the RPE, carrying out suspension cultivation to obtain 3D analogous retina comprising NR tissues, then carrying out continuous suspension cultivation inside an optimized culture solution without retinoic acid, carrying out NR differentiation on all retinal cells, comprising highly matured photoreceptor cells, Rhodopsin positive rod cells, L / M OPSIN positive red and green cone cells and S-OPSIN positive blue cone cells, and thus particularly obtaining the analogous retinal tissues rich in red and green cone and rod cells.