996 results about "Culture vessel" patented technology

A novel growing method is provided for pluripotent stem cells such as ES cells. The method of the invention is a pluripotent stem cell growing method and gene transfer method in which pluripotent stem cells are cultured under conditions that maintain their undifferentiated state and pluripotency, the method being characterized by using a liquid medium and a culturing vessel having immobilized or coated on a substrate solid phase surface a molecule which is adhesive to the pluripotent stem cells in a fixed concentration, to grow the pluripotent stem cells in a dispersed state while maintaining their undifferentiated state and pluripotency, without using feeder cells, or to transfer and express a gene therein.

The present invention provides a culture vessel and microscope that allows a wide area of favorable observation of the sample in the plural wells even when a negative refractive power is generated in the culture medium, wherein a ring-shaped opening, a condenser lens, a lens array, a well plate, an objective lens and a phase contrast plate are disposed in this order viewed from the light source side with a construction to place a phase contrast film of the phase contrast plate in a conjugated relation to the projection pupil position or projection pupil of the objective lens, and the culture vessel is constructed of the lens array and well plate with a relative disposition in which the optical axis of each lens is coaxial with the center line of the confronting each well; the aperture of the ring-shaped opening being smaller than the aperture of the ring-shaped opening that is used when the culture vessel is not disposed in the optical path.

The invention provides a method for large-scale production of a polypeptide, such as a Factor VII or Factor VIIa polypeptide, in eukaryote cells, such as mammalian cells, contained in a culture liquid, said method comprising: monitoring the concentration of dissolved CO2 in the culture liquid, and constantly or intermittently sparging atmospheric air through the culture liquid, wherein the sparging rate of the air is controlled in relation to the monitored concentration of dissolved CO2 in the culture liquid. The method reduces or eliminates the use of bases while providing an excellent pH control. The invention also provides a culture vessel suitable for the methods.

The present invention provides methods for generating mammalian T cell populations comprising antigen-specific CD4+CD25+ regulatory T cells from freshly isolated CD4+CD25− T cells. The method comprises selecting CD4+CD25− T cells from a sample obtained from a mammalian subject; determining the MHC Class II type of the subject; inducing the generation of antigen-specific regulatory T cells by contacting the isolated CD4+CD25− T cells in a culture vessel with an induction agent for a time period sufficient to generate antigen-specific CD4+CD25+ regulatory T cells; and selecting the CD4+CD25+ antigen-specific regulatory T cells by sorting the cells in the induction culture with a selection agent comprising at least one artificial multimeric MHC Class II / peptide complex that corresponds to the MHC Class II type of the subject.

The present invention provides a biological sample culturing and observation system which includes: an incubator in which a biological samples are housed, and whose interior is maintained in a culturing environment satisfying predetermined conditions, and the interior is isolated from the outside; an observation optical system that from outside the incubator optically observes the biological sample via the incubator; a light blocking device that blocks external light that is irradiated on the biological sample and within the visual field of a observation optical system; and a supply device that selectively supplies a liquid or gas to the biological sample inside the incubator from a plurality of holding vessels that individually hold a plurality of different types of liquid or gas that are necessary for the culturing.

The present invention provides a cell culture data and cell image capturing and remote monitoring system that includes an imaging sensor pod having one or more cameras to capture multiple color still and / or motion images of cell cultures and cells located on a support and / or in a liquid located in a culture vessel within an incubator. The images can be taken at varying magnifications, and from different discrete locations on the support or in the vessel round the clock. The captured data and images are wirelessly transmitted to a management control unit, and / or another wirelessly connected and remotely located data transmission device such as PDA, for a researcher to review and analyze to determine the health and viability of cells, and the status of the cell culture. The image capturing and remote monitoring system taught herein alleviates the need for a researcher to be in the physical presence of a biological sample in order to visually ascertain and analyze the health and viability of the sample and status of the cell culture.

The present invention provides a variety of microscale bioreactors (microfermentors) and microscale bioreactor arrays for use in culturing cells. The microfermentors include a vessel for culturing cells and means for providing oxygen to the interior of the vessel at a concentration sufficient to support cell growth, e.g., growth of bacterial cells. Depending on the embodiment, the microfermentor vessel may have various interior volumes less than approximately 1 ml. The microfermentors may include an aeration membrane and optionally a variety of sensing devices. The invention further provides a chamber to contain the microfermentors and microfermentor arrays and to provide environmental control. Certain of the microfermentors include a second chamber that may be used, e.g., to provide oxygen, nutrients, pH control, etc., to the culture vessel and / or to remove metabolites, etc. Various methods of using the microfermentors, e.g., to select optimum cell strains or bioprocess parameters are provided. The invention provides microreactors having a variety of different designs, some of which incorporate active stirring and / or have the capability to operate in batch or fed-batch mode. The invention further provides an apparatus and methods for simultaneous operation of a plurality of microreactors, with monitoring of the individual microreactors during a run. The invention further provides methods of performing gene expression analysis on cells cultured in microreactors.

The invention provides an automated apparatus of cell culture having tanks of culture medium, of growth factors and of cells to be cultured, an incubator having a thermostated enclosure which houses a cell culture vessel, and control computer system. A supporting and agitation device of the culture vessel is provided in the enclosure, and the culture vessel is formed by a bag having at least one inlet port connected to the tanks and one outlet port connected devices for harvesting and storage of the cells after culture, these harvesting and storage devices and tanks being located outside the enclosure and being connected to the cell expansion bag ports by conduits which together with the cell expansion bag form a preassembled module passing through a wall of the enclosure.

The present invention provides methods for generating mammalian T cell populations comprising antigen-specific CD4+CD25+ regulatory T cells from freshly isolated CD4+CD25− T cells. The method comprises selecting CD4+CD25− T cells from a sample obtained from a mammalian subject; determining the MHC Class II type of the subject; inducing the generation of antigen-specific regulatory T cells by contacting the isolated CD4+CD25− T cells in a culture vessel with an induction agent for a time period sufficient to generate antigen-specific CD4+CD25+ regulatory T cells; and selecting the CD4+CD25+ antigen-specific regulatory T cells by sorting the cells in the induction culture with a selection agent comprising at least one artificial multimeric MHC Class II / peptide complex that corresponds to the MHC Class II type of the subject.

A microscope stage capable of always heating the entire culture vessel even when the culture vessel is moved in two-dimensional directions, and allowing an object lens and a condenser even in a high-power microscope to approach an object of observation such as a cell until they come into focus. A heating unit (58) faces a well plate (37) on a drive base (49) even when the drive base (49) is driven to any position in two-dimensional directions. Therefore, all the cells A in the small compartments (45) of the well plate (37) are always heated. Since the microscope stage (25) is constituted such that a lower-side base (71) houses an upper-side base (73) and a fixed base (47) in recessed portions, its thickness is considerably small. Accordingly, an object lens (5) and a condenser (3) can approach close to the cells A. Therefore, it is possible for high-power object lens (5) and condenser (3) to focus on the cells A.

The invention intends to provide a cell culture apparatus which is able to realize an adequate culture according to the culture state of cells while alleviating the labor of an operator. The cell culture apparatus includes a culture bag for causing the cells to proliferate, a cell inoculation cassette (or culture bag as an antibody stimulating and proliferation culture vessel) for stimulating the cells by an inducer for the proliferation, a culture medium cassette for storing a culture medium supplied to the culture bag and the cell inoculation cassette, a CCD camera 88 for acquiring images of the cells in the cell inoculation cassette, and an image processing computer and an operation control computer for determining the culture state (proliferation capability and proliferation ability of the cells) of the cells from the images of the cells acquired by the CCD camera, and causing a culture operation to be carried out on the basis of the determination.

The invention relates to a method and device for preparing functional nanoparticle / bacterial cellulose composite membranes by an inverted culturing method. The method comprises the steps of preparing a culture medium, carrying out activation and accessing on strains, and carrying out culturing and post-processing on bacterial cellulose membranes, wherein in the step of preparing the culture medium, functional nanoparticles are added. Inverted culturing refers to inversely place a culturing vessel on an oxygen chamber, the culturing vessel and the oxygen chamber are separated by using an oxygen permeating membrane, and a seam between the culturing vessel and the oxygen chamber is sealed. The device disclosed by the invention comprises a base and a culturing vessel, wherein the base is an open vessel with a step at the upper part thereof, the culturing vessel is inversely placed on the step of the vessel, the base and the culturing vessel are separated by using an oxygen permeating membrane, the oxygen permeating membrane and the lower part of the base form an oxygen chamber, and the lower part of the base is provided with an air outlet hole and an air inlet hole. The method and device disclosed by the invention integrate the culturing and functionalization of bacterial cellulose membranes, and are simple in operation, short in cycle and low in cost; and according to the invention, on the premise of not damaging a three-dimensional network structure in a bacterial cellulose membrane, a functionalized bacterial cellulose composite membrane which is good in functional-nanoparticle adsorption property and excellent in performance is prepared.

There is provided a technology for simple three-dimensional culturing of cartilage cells. The technology includes a method for culturing cartilage cells which comprises seeding cartilage cells 1 on a plurality of projected portions 21 of a culturing substrate 2 having a culture surface on which the projected portions 21 having an equivalent diameter and an interval which are smaller than the equivalent diameter of the cartilage cells 1 to be cultured, placing the culture substrate 2 having the cartilage cells 1 in a culture vessel, and culturing the cartilage cells to form aggregates of cartilage cells.

The invention provides a method for preparing a fractal structure microchannel by 3D printing, comprising: 1) drawing by computer drawing software, and printing out a corresponding fractal structure skeleton by a 3D printer; 2) fixing the fractal structure skeleton on a culture dish , and inject polydimethylsiloxane solution to a certain height; 3) vacuum the air bubbles in the polydimethylsiloxane solution, heat and solidify; 4) use a solvent to dissolve the fractal structure skeleton to form a fractal Structural microchannels. The preparation process of the present invention is simple, does not require complex instruments and strict laboratory conditions, can multi-layer complex structure microchannels and does not need to assemble multiple parts, and is especially suitable for fast, low-cost, large-scale complex structure channels in microfluidic technology. Batch production.

The invention relates to a bioreactor and method, particularly a reactor and method for seed cell culture or cell / biological material compound culture in cytobiology, tissue engineering and medicine. The method for biological culture of cell or tissue engineering comprises the following steps: putting biological culture raw materials of cell or tissue engineering into a culture vessel with a U-shaped bottom surface, W-shaped bottom surface or V-shaped bottom surface; and driving the culture vessel to annularly swing by using an annular swing power system, wherein the culture vessel annularly swings by using the center as the pivot, and each point on the same circumference sequentially swings up and down. The invention also discloses a device for implementing the method. Under the action of the power provided by the external annular swing, the invention can reduce the cell damage and provide a uniform culture environment under ideal mass transfer conditions.

The invention discloses an apparatus for growth of a cell colony. The apparatus provided by the invention comprises a micro graphical pattern template with a plurality of micro graphical cavities and a viscous hydrogel disposed under the micro graphical pattern template; the micro graphical pattern template and the viscous hydrogel are cross-linked and adhered into a whole; the shape and size of each micro graphical cavity determines a physical space of the growth of the cell colony. Materials used for the viscous hydrogel can be composed of a natural biomaterial and / or an artificially synthesized biomaterial capable of cross-linkedly forming the hydrogel and a cross-linking agent. A platform provided by the invention is simple in preparation process, is simple and practical for use matching with a culture dish. A plurality of co-culture environments are constructed by using the micro graphical technology, so that convenient and practical means are provided for construction and screening of high-throughput model; at the same time, independent control a plurality of factors such as the shape and size of the cell colony, mechanical strength and chemical components of an adhesive substrate, growth factors and chemical signals, co-culture cells and the like; and cell proliferation culture and induced differentiation culture can be carried out in-situ.

The invention discloses a method for producing Anrodia camphorata mycelia based on solid-state surface culture, and belongs to the field of bioengineering. The method comprises the following steps of: screening components of a culture medium of the Anrodia camphorate and optimizing the proportion of the cereal raw material in the culture medium so that the culture medium is firmly adsorbed to the inner wall of a culture vessel or the culture surface to bring convenience to the surface culture of the Anrodia camphorata mycelia; inoculating the Anrodia camphorata mycelia in the culture surface of a reactor and culturing the Anrodia camphorata mycelia in the environment of certain temperature and moisture; feeding the culture medium every a certain time in batch during the surface culture so as to timely replenish the nutrient components required by the growth of the Anrodia camphorata mycelia and making the growth environment of the Anrodia camphorata mycelia close to the natural culture environment until a large amount of mycelia or large colony is formed; and quickly drying the Anrodia camphorata culture at a low temperature after the culture. The method of the invention makes the growth environment of the Anrodia camphorata mycelia close to that of the wild Anrodia camphorate and improves the productivity of the Anrodia camphorata culture by timely feeding the culture medium in batch, so that the obtained Anrodia camphorata culture has high content of active ingredient and closer efficacy to the wild fruit body.

The present invention provides devices and methods for achieving mixing and gas exchange in chambers having a small volume. According to the invention a chamber includes one or more walls that are made of a gas-permeable material and that include multiple portions that are selectively deflectable into the interior of the chamber. The deflectable portions are in communication with a hollow cavity or space that can be pressurized. Pressurization results in deflection of the deflectable portion into the chamber. Pressurization is achieved using a gas or fluid that contains a gas of interest such as oxygen. The portions are deflected in a sequence that results in peristaltic action that mixes and oxygenates the contents of the chamber. In a preferred embodiment of the invention the hollow cavities are an assembly of tubes that deflect into a plurality of chambers. A particular use for the invention is as a culture vessel for cells such as bacteria.

There are provided a culture treatment apparatus and an automatic culture apparatus capable of taking quick and appropriate action even if there is a change in the state of a space where cells stored in a container, such as a culture vessel, for the automatic culture apparatus are subjected to predetermined treatment. The culture treatment apparatus includes a treatment section for applying predetermined treatment to cells stored in an openable container for the automatic culture apparatus in a partitioned space; a detection section for detecting a predetermined state of the space, the detection section being in the space or in the vicinity of the space; and a control section for controlling the treatment section to close the container if the detection section detects the predetermined state with the container opened.

A culturing apparatus has a culture vessel having a first elastic seal bonded on its upper face and a culture space formed thereon, and a joint having supplying means for supplying solution such as a medium to this culture vessel and a second elastic seal having microprojection formed at the lower face. The first elastic seal has a valve for supplying or discharging solution. The second elastic seal is formed with microprojection at the position corresponding to a valve for preventing a spill. The culture vessel and the joint are sucked by the adsorbing means between the first elastic seal and the second elastic seal, thereby forming an integral seeding device.

The invention provides a full automatic microorganism sampler, which can solve the problems that specially-assigned persons are needed and only single sampling is carried out in the prior art. The technical scheme is achieved as follows: the full automatic microorganism sampler comprises a shell part, a circuit part, a gas circuit part, a vessel storing part consisting of a sterilized vessel storing part and a bacterial vessel storing part, a sampling head part consisting of a movable sampling head part and a fixed sampling head part and a manipulator part, wherein the manipulator part clampsa culture vessel and moves among the fixed sampling head part, the sterilized vessel storing part and the bacterial vessel storing part; the sampling head part, the manipulator part and the vessel storing part are all arranged on a bottom board of the shell part; and the sampler adopts a GPRS wireless data transmission technology. Compared with the prior sampler, the full automatic microorganism sampler has the advantage of high automation degree and realizes unattended automatic vessel replacement, automatic continuous sampling and automatic recording of sampling data.

A method of growing chondrocytes on an allograft cancellous bone structure which had been previously treated by demineralization to form a cartilage repair implant comprises isolating allograft chondrocytes from articular cartilage of a donor other than the patient on which the implant is to be used, cultivating the isolated chondrocytes in a medium to obtain a dedifferentiated fibroblast-like phenotype, placing an at least partially demineralized allograft cancellous bone structure carrier having at least one surface prepared for cell seeding in a culture vessel; adding the cultivated dedifferentiated chondrocytes to a demineralized surface of the cancellous bone structure carrier, and incubating the cancerous bone structure carrier for a period of time of at least 40 days to obtain an implant with a cartilage layer which ranges from about 2 mm to 5 mm n thickness.

The invention intends to provide a cell culture apparatus which is able to realize an adequate culture according to the culture state of cells while alleviating the labor of an operator. The cell culture apparatus includes a culture bag for causing the cells to proliferate, a cell inoculation cassette (or culture bag as an antibody stimulating and proliferation culture vessel) for stimulating the cells by an inducer for the proliferation, a culture medium cassette for storing a culture medium supplied to the culture bag and the cell inoculation cassette, a CCD camera 88 for acquiring images of the cells in the cell inoculation cassette, and an image processing computer and an operation control computer for determining the culture state (proliferation capability and proliferation ability of the cells) of the cells from the images of the cells acquired by the CCD camera, and causing a culture operation to be carried out on the basis of the determination.

A method for culturing cells in a plastic culture vessel is provided. The method is suitable for use when it is desirable to have a small hydrophobic molecule present in the cell culture media at some point during incubation. A plastic cell culture vessel is also provided which is made of a light blocking material capable of blocking exposure of the biological fluid within from light.

The invention relates to a method for rapidly cultivating chlorella and belongs to the technical field of biology. The cultivation method comprises the following steps: preparing a standard BG-11 culture medium to serve as a basal culture medium of chlorella by taking fresh running water as a solvent; adding a certain amount of organic carbon source and NaHCO3 into the standard culture medium, so that the final concentration is respectively 0.5-1.0g / L and 0.2-0.5g / L; putting the culture medium into a culture vessel, inoculating 10-20 percent of chlorella solution, sealing the opening, and culturing the chlorella solution in an environment with the illumination intensity of 3000-4000Lx, the light-dark ratio of 12h:12h and the temperature of 22-28 DEG C. According to the method, the cell density of the chlorella can reach a high level in a short cultivating cycle and is increased by about six times compared with that cultivated by using the basal culture medium. Meanwhile, the method has the advantages that the process is easy to operate, the culture medium is not required to be sterilized, the pH value of the culture medium is not required to be regulated, and the method can be widely applied to cultivating chlorella in industrial production.

The invention discloses a method for preparing a population of?human pluripotent stem cells and the application thereof. The preparation of stem cells is characterized by comprising the following steps: CD151<+>, CD31<->, Sox<2+> pluripotent stem cells are separated and collected from human umbilical cord and or placenta tissues; the cells adhere to grow in a culture vessel under a predetermined condition and expand through passage 20 or above to be still stable in gene expression. The population of cells of this invention do not form teratoma after injection into animals. The human pluripotent stem cells highly express CD151, OCT4 and Sox-2 as specific markers of embryonic stem cells, as well as specific markers of epidermic cells, endothelial cells, thrombocytes, dendritic cells, while lack expression of CD31, CD34, CD45 and HLA-II. The pluripotent stem cells are also characterized as being able to adhere to tissue culture plastic and having the potential to differentiate into three germ layers: endoderm, mesoderm and ectoderm. These pluripotent stem cells are able to be used as carrier cells of gene therapy and for the treatment of diseases caused by cell damage or cell aging. The present invention provides a method of isolating, purifying and culturally expanding of a population of human pluripotent stem cell for preparing the high purity injection preparation. The preparation of stem cells has a good therapeutic effect on the treatment of diseases caused by cell damage or cell aging in animal and human clinical trials. The preparation also has no toxic side effect and no immune rejection.

There is provided a cell culture evaluation system, a cell culture evaluation method, and a cell culture evaluation program which are capable of estimating and evaluating the lag time or the minimum doubling time and objectively and adequately determining whether or not a cell population is stimulated for proliferation by using an average projected area of a cultured cell population or the rate of increasing the ratio of the non-single-cells as an evaluation parameter when culturing the cells. Images of the cell population to be cultured statically are acquired in a culture vessel, the average projected areas of the cells are calculated from the images for the respective culture times, and the lag times at lag phase are calculated from the calculated average projected areas of the cells. The single-cells and the non-single-cells are discriminated from the images, and the increasing rate of the non-single-cells is calculated from the ratio of the non-single-cells in the cell population to determine the minimum doubling time from the increasing rate of the non-single-cells. Whether or not the cell population is stimulated for proliferation is determined from the ratio of the non-single-cells.

The present invention provides a culture vessel for forming an aggregated cell mass which has at least two wells, wherein the wells are made from a light-shielding member and an inner surface of the wells is subjected to treatment to impart cellular non-adhesiveness.

A culture-vessel adaptor that is detachably fixed to the exterior of a culture vessel capable of holding a culture medium containing a specimen is provided. The culture-vessel adaptor includes a vessel mounting part where the culture vessel is mounted; and a protruding part disposed so as to extend outward from a side surface of the culture vessel when the culture vessel in mounted in the vessel mounting part. Handling of culture vessels becomes easy by an automated mechanism such as a robot, with a straightforward and low-cost method, and the culture vessels can be reliably prevented from being dropped.