589 results about "Adherent cell" patented technology

A multi-layer, flexible, gas-permeable film (10) suitable for forming a cell culture container (20), the film (10) comprising a first layer (12) composed of a polystyrene having a thickness within the range of 0.0001 inches to about 0.0010 inches and, a second layer (14) adhered to the first layer (12) composed of a polyolefin having a thickness within the range of 0.004 inches to about 0.015 inches.

We introduce a new method for implementing cell-based assays and long-term cell culture. The method is based on digital microfluidics (DMF) which is used to actuate nanoliter droplets of reagents and cells on a planar array of electrodes. DMF method is sutable for assaying and culturing both cells in suspension and cells grown on surface (adherent cells). This method is advantageous for cell culture and assays due to the automated manipulation of multiple reagents in addition to reduced reagent use and analysis time. No adverse effects of actuation by DMF were observed in assays for cell viability, proliferation, and biochemistry. These results suggest that DMF has great potential as a simple yet versatile analytical tool for implementing cell-based assays and cell culture on the microscale.

Devices and methods for implementing cell-based assays and long-term cell culture. The device and method are based on digital microfluidics (DMF) which is used to actuate nanoliter droplets of reagents and cells on a planar array of electrodes. DMF method is suitable for assaying and culturing both cells in suspension and cells grown on surface (adherent cells). This method is advantageous for cell culture and assays due to the automated manipulation of multiple reagents in addition to reduced reagent use and analysis time. No adverse effects of actuation by DMF were observed in assays for cell viability, proliferation, and biochemistry. These results suggest that DMF has great potential as a simple yet versatile analytical tool for implementing cell-based assays and cell culture on the microscale.

The use of an imaging system, cell compartment markers, and molecular markers in methods for correlating the movement of molecules within a cell to a particular compartment are provided, including measuring and correlating molecule movement in adherent and non-adherent cells.

In biology and biotechnology, electroporation is an important technique for introducing entities (DNA, RNAi, peptides, proteins, antibodies, genes, small molecules, nanoparticles, etc.) into cells. Applications range widely from genetic engineering to regenerative medicine to drug delivery. It has been demonstrated that the electrical currents flowing through cells can be used to monitor and control the process of electroporation for biological and artificial cells. In this application, a device and system are disclosed which allow precise monitoring and controlling electroporation of cells and cell layers, with examples shown using adherent cells grown on porous membranes.

Systems and methods are providing for performing high-throughput, programmable, multiplexed assays of biological, chemical or biochemical systems. Preferably, a micro-pallet includes a small flat surface designed for single adherent cells to plate, a cell plating region designed to protect the cells, and shaping designed to enable or improve flow-through operation. The micro-pallet is preferably patterned in a readily identifiable manner and sized to accommodate a single cell to which it is comparable in size. Each cell thus has its own mobile surface. The cell can be transported from place to place and be directed into a system similar to a flow cytometer. Since, since the surface itself may be tagged (e.g., a bar code), multiple cells of different origin and history may be placed into the same experiment allowing multiplexed experiments to be performed.

A method of treating ischemia in a subject in need thereof is disclosed. The method comprising administering to the subject a therapeutically effective amount of adherent cells of a tissue selected from the group consisting of a placenta and an adipose tissue, thereby treating the ischemia in the subject. A method of treating a medical condition requiring connective tissue regeneration and / or repair is also disclosed.

The present invention relates to treatment of polymeric bioreactor surfaces, to promote the proliferation of adherent cells.

A method of cell expansion is provided. The method comprising culturing adherent cells from placenta or adipose tissue under three-dimensional culturing conditions, which support cell expansion.

An apparatus for collecting or culturing cells or cell colonies includes: a common substrate formed from a flexible resilient polymeric material and having a plurality of wells formed therein; and a plurality of rigid cell carriers releasably connected to said common substrate, with said carriers arranged in the form of an array, and with each of the carriers resiliently received in one of the wells. A method of collecting or culturing cells or cell colonies with such an apparatus is carried out by depositing a liquid media carrying cells on the apparatus so that said cells settle on or adhere to said the carriers; and then (c) releasing at least one selected carrier having said cells thereon by gradual application of release energy to each carrier from the cavity in which it is received (e.g., by pushing with a probe).

Provided are a method capable of evaluating adherent cells under an environment similar to an in vivo environment by a culture method similar to a two-dimensional culture, and applications thereof. An adherent cell culture method uses, as a culture chamber (10), a chamber in which two or more culture spaces each having an equivalent diameter (D) that is 1 to 5 times the diameter of a desired spheroid and each having a height (H) that is 0.3 to 5 times the equivalent diameter are arranged and a surface of each of the culture spaces has a water contact angle of 45 degrees or less. Spheroids of adherent cells are cultured in the respective culture spaces (11) arranged in the culture chamber (10).

An improved prosthetic graft for the bypass, replacement or repair of vessels and organs that are in contact with blood flow is disclosed. The prosthetic graft includes a porous prosthetic implant and adherent cells adhered to the outer surface of the implant. The adherent cells are transfected with at least one recombinant nucleic acid molecule encoding at least one protein that enhances patency of the graft. The prosthetic graft has a long-term patency and success rate that is superior to other previously described prosthetic grafts designed for such use. Also disclosed are methods of making and using such a graft.

The present invention provides a microorganism culture array, including one or more microflow cell culture array units, the microflow cell culture array unit includes a main channel and a plurality of cell culture units connected with the main channel, each culture unit includes a diffusion channel and a cell culture room, the cell culture room is connected with the main channel. The microorganism culture array of the invention not only realizes isolation of single-cell, but also makes the cell locate in zero flow state when the culture liquid or other reagent is changed, bringing greatly convenience for the cell culture and observation especially the culture and observation of non-adherent cells, greatly fit for research work of high throughput screen based on the cell phenotype.

A method of treating ulcerative colitis or Crohn's disease in a subject in need thereof is disclosed. The method comprising administering to the subject a therapeutically effective amount of adherent cells from a placenta or adipose tissue, thereby treating the ulcerative colitis or Crohn's disease.

The invention discloses an efficient multiplication CTL preparation method killing tumors in a targeted mode. The CTL preparation method comprises the following steps: (a) removing CD4+CD25+Treg cells through immunomagnetic bead negative sorting; (b) arranging mixed cells in a serum-free medium for cultivation, and obtaining suspension cells and adherent cells; (c) adding GM-SCF and IL-4 in the adherent cells, culturing the cells for five days; in the sixth day, adding a tumour cell holoantigen, and in the seventh day, adding TNF-alpha and IL-27; (d) transferring the suspension cells to a culture flask wrapped by a CD3 monoclonal antibody and recombinant human fibronectin, adding IFN-gamma, in the second day, adding IL-2, IL-12 and the IL-27, and culturing the mixture till the eighth day to obtain CIK cells; (e) mixing the CIK cells and mature DC cells, and adding the IL-12, IL-7 and an anti-CD 28 monoclonal antibody for cultivation; in the third day, adding an anti-CTLA-4 monoclonal antibody, and then culturing the mixture for four days. According to the efficient multiplication CTL preparation method killing tumors in the targeted mode, efficiency of in-vitro CTL cell proliferation is improved, activity of killing the tumor cells in the targeted mode is improved, transformation of peripheral blood mononuclear cells to the CD4+CD25+Treg cells is inhibited.

The invention provides an inducing method and inducing culture medium for differentiation of bone marrow mesenchymal stem cells into osteoblasts in vitro. The inducing culture medium is composed of 1*10<-8> mol / L of dexamethasone, 50 mu mol / L of ascorbic acid and 10 mmol / L of sodium beta-glycerophosphate; and solvent is a supernatant of a sclerite complete culture medium and comprises 10% of fetal calf serum, 100 U / mL of penicillin, 100 mg / L of streptomycin, a mixture of DMEM culture fluid and F12 culture fluid and multiple growth factors secreted by bone cells in the sclerite culture process. According to the invention, bone marrow mesenchymal stem cells of a mouse are purified by replacing the cell culture fluid through an adherent cell passage method, the obtained cells of the first generation are induced, and the supernatant of the sclerite complete culture medium cultured for 72-96 hours is used as the solvent of osteoblast differentiation inducer, thereby obviously improving the in vitro osteogenic differentiation efficiency of bone marrow mesenchymal stem cells.

An in vitro biomechanical model used to applied hemodynamic (i.e., blood flow) patterns modeled after the human circulation to human / animal cells in culture. This model replicates hemodynamic flow patterns that are measured directly from the human circulation using non-invasive magnetic resonance imaging and translated to the motor that controls the rotation of the cone. The cone is submerged in fluid (i.e., cell culture media) and brought into close proximity to the surface of the cells that are grown on the plate surface. The rotation of the cone transduces momentum on the fluid and creates time-varying shear stresses on the plate or cellular surface. This model most closely mimics the physiological hemodynamic forces imparted on endothelial cells (cell lining blood vessels) in vivo and overcomes previous flow devices limited in applying more simplified nonphysiological flow patterns. Another aspect of this invention is directed to incorporate a transwell co-cultured dish. This permits two to three or more different cell types to be physically separated within the culture dish environment, while the inner cellular surface is exposed to the simulated hemodynamic flow patterns. Other significant modifications include custom in-flow and out-flow tubing to supply media, drugs, etc. separately and independently to both the inner and outer chambers of the coculture model. External components are used to control for physiological temperature and gas concentration. The physical separation of adherent cells by the artificial transwell membrane and the bottom of the Petri dish permits each cell layer, or surface to be separately isolated for an array of biological analyses (i.e., protein, gene, etc.).

A method of selecting a population of adherent cells of a placenta tissue suitable for transplantation is disclosed. The method comprising: (a) determining prior to transplantation in a candidate population of adherent cells of a placenta tissue at least one of the following parameters: (i) percentage of viable cells in the candidate population; (ii) immune phenotype of cells in the candidate population; (iii) xeno-contamination in the candidate population; (iv) sterility of the candidate population; and (v) immunosuppressive activity of cells in the candidate population; and (b) selecting or excluding the candidate population according to predetermined values of at least one of the parameters, thereby selecting a population of adherent cells of the placenta tissue suitable for transplantation.

The invention discloses a method for primary culture of tumor cells and relates to the field of cell biology. According to the method disclosed by the invention, a tumor sample is prepared into a single-cell suspension liquid by use of an enzyme method or a mechanical method, and then tumor cell bladders are prepared by use of a serum-free suspension culture method, wherein alkaline fibroblast growth factors, epidermal growth factors, insulin and bovine serum albumin need to be added during a culturing process, then the growth factors are removed, and the culture is implemented in a general culture medium to obtain adherent cells capable of realizing continuous passage. The method disclosed by the invention is short in culture period and has the effect of increasing the success rate of the primary culture of tumors.

A method of harvesting mesenchymal stem cells from human amniotic fluid uses a two-stage culture protocol comprising culturing human amniocytes and then culturing mesenchymal stem cells. For culturing human amniocytes, primary amniocyte cultures are set up using routine or standard culture protocol in a cytogenetic laboratory. Non-adherent human amniotic fluids cells in the supernatant medium are collected. For culturing mesenchymal stem cells (“MSC”), the non-adherent cells are centrifuged and then plated with an alpha-modified Minimum Essential Medium supplemented with fetal bovine serum. Incubate with humidified CO2 for MSC growth. Reverse transcription polymerase chain reaction (“RT-PCR”) and immunocytochemical analyses reveal that Oct-4 mRNA and OCT-4 protein expression is detectable in the cultured amniotic fluid mesenchymal stem cells (“AFMSCs”). Under differentiation culture conditions, the AFMSCs can be induced to develop into multi-lineage cells, such as adipocytes, osteocytes, neuronal cells, etc.

An electroporation apparatus provides for the electroporation of adherent cells attached to an electrode surface or suspended cells in close proximity to an electrode surface. In one embodiment, the electrodes are transparent to allow cell viewing using a microscope or an automated image analysis machine. The geometry of the electrodes and associated electrically non-conductive structures may provide for well-defined regions of electroporated and non-electroporated adherent cells with a clearly defined interface between these regions, facilitating comparison of electroporated cells and non-electroporated cells, and evaluation of transfer of material from cell to cell via intercellular gap junctions.

The invention relates to a device and method for noninvasive continuous monitoring of quantity or concentration of dynamic cells, in particular to a device and method for noninvasive continuous on-line or off-line monitoring of quantity or concentration of dynamic cells in adherent cell cultures (usually as stem cell cultures or other therapeutic cell cultures) or cell and tissue cultures. The device comprises: a cell culture medium supply system, one / one type of or multiple / multiple types of upstream biomedical indicator detection head(s) or connector(s), a cell culture , one / one type of or multiple / multiple types of downstream biomedical indicator detection head(s) or connector(s), one or multiple speed controllable sterile driving unit(s) of fluid, a waste or harvested liquor system, liquid conveying pipeline systems that are driven and controlled in speed by the speed controllable sterile driving unit(s) of fluid for connecting the above components in order. And the upstream and downstream biomedical indicator detection head(s) or connector(s) are connected to a monitoring and controlling system and / or a computer so as to obtain, process and monitor data, and furthermore provide feedbacks for the whole cell culture system through a signal feedback system for realizing control.

A carrier for growing adherent cells is provided, wherein the carrier comprises one or more outer surfaces; and one or more structured indentations on one or more of the outer surfaces, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. The carrier may comprise a single indentation or ‘cup’ like structure, or may comprise a plurality of indentations. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.

The invention provides a stem cell culture medium and a method for culturing endometrium stem cells by using the stem cell culture medium. The method comprises the following steps: separately collecting menstrual blood and endometrium tissues, respectively culturing the menstrual blood and endometrium tissues in the stem cell culture medium provided by the invention to respectively obtain menstrual blood adherent cells and endometrium adherent cells, culturing the menstrual blood adherent cells and endometrium adherent cells in a cell culture bottle, collecting the adherent cells by trypsinization, inoculating the adherent cells in a cell coculture dish, and culturing the adherent cells in the stem cell culture medium provided by the invention. The stem cell culture medium has the advantages of simple components, fewer added components and lower cost. After more than 20 generations of in-vitro culture, the cells can not easily have the phenomenon of aging or degeneration, and can maintain the activity and stem property of the stem cells for a long time. The stem cell culture method is simple and effective, the cell proliferation efficiency is high, and the in-vitro culture doubling time is only 20 hours or so. The cells can be stably amplified by 50 generations.

Adherent cells and other membranous structures that are immobilized on a solid surface are transfected by electroporation in which the electric field is produced by a array of closely spaced electrodes positioned above the surface. Each electrode is substantially smaller in at least one lateral dimension than the dimensions of a single cell, and the electrodes in each pair are spaced apart by distances selected such that that a maximum of one cell will reside within the field produced by each pair, and the distance of the electrodes above the surface to which the cells are adherent is small enough to place the cell within the resulting electric field and yet great enough to avoid contact of the electrodes with the cell membrane.

A method for isolating and processing bone marrow derived stem cells, including the steps of: (a) collecting a biological sample containing adherent bone marrow stem cells in a receptacle with interior walls coated with a cell-adherent substrate; (b) incubating the bone marrow cells on the adherent substrate so that a layer of adherent bone marrow stem cells adheres to the substrate; (c) washing any non-adherent cells from the substrate; and (d) collecting the bone marrow stem cell layer. Isolation kits and use of bone marrow cells harvested for cell therapies are also described.

The invention relates to the field of cells, particularly to a cell cryoprotectant and application thereof. The cell cryoprotectant is composed of mixtures of permeable protectant, non-permeable protectant, energy supply agent and the like and can be applied to cryopreservation of adherent cells, suspension cell and clinic application cells. The survival rate of cells recovered and cultured in thecell cryoprotectant can reach up to 98%, and detected various indexes of the cells show that the cells are functionally heathy, so that the cell cryoprotectant can well maintain the physiological functions of the cells and avoid mutation, tumorigenicity and the like. Meanwhile, the cell cryoprotectant is simple in preparation processes, easy to operate, wide in application range and capable of facilitating research and clinical popularization and application.

The present invention relates to the field of adherent cell culture. More closely, the invention relates to a method for production of a cell attachment and culture surface, such as a microcarrier, comprising a guanidino-containing ligand, wherein the ligand is coupled via reaction involving a primary amine to the surface which is activated by activation groups such that the final molar ratio of grafted ligand and ungrafted activation groups is above 1.5. Preferably, the ligand density is above 0.5 mmol / g cell culture surface and the remaining activation groups after coupling is less than 0.6 mmol / g cell culture surface. The cell culture surface may be used for various purposes, primarily cell cultivation and virus production.