Abstract: An apparatus and method is described for seeding and culturing cells on a sample. The apparatus includes a chamber in which the volume of the chamber may be adjusted without compromising the seal or sterility of the chamber. The apparatus enables the seeding of cells in a reduced volume and culturing of cells in an increased volume. Further, the apparatus enables application of forces, strains and torques to a sample during seeding, culturing or transportation of the sample.

Abstract: A versatile compartmentalized cell culture device, with a selectively permeable membrane separating the compartments, provides many attributes relative to traditional devices. It can be configured for high-density cell culture, co-culture, and sample dialysis while rolling or standing still. It can also be configured for continuous movement of liquid between compartments. The wide combination of attributes not found in other membrane based cell culture and bioprocessing devices includes more cell capacity, more cell secreted product capacity, higher cell and product density, increased medium capacity, minimized use of exogenous growth factors, compatibility with standard cell culture equipment and protocols, increased scale up efficiency, capacity to function when rolling or standing still, capacity for perfusion without the need for pumps, and more efficient sample dialysis.

Abstract: The present invention refers to a method of fabricating a membrane made of a nanostructured material and its use.

Abstract: Described are preferred extracellular matrix composites including a first extracellular matrix material having a second extracellular matrix material deposited thereon. The preferred materials are made by culturing cells in contact with an extracellular matrix graft material in a fashion to cause the cells to biosynthesize and deposit extracellular matrix components on the material. The cells are then removed to provide the extracellular matrix composite material.

Abstract: Disclosed is a microfluidic chip and method using the same. The microfluidic chip comprises a substrate having a surface, and at least a tissue culture area formed on the surface of the substrate. The tissue culture area has a microfluidic channel formed by a plurality of connected geometrical structures (nozzle-type channels) having a predetermined depth. The microfluidic channel has an inlet and an outlet, which are at two ends of the microfluidic channel, for medium inputting and outputting, respectively. Additionally, at least an air-exchange hole is formed on the bottom of the microfluidic channel. By using the microfluidic chip for tissue culture, lateral flow speed and stress can be decreased, so as to prolong survival time of tissues (e.g. liver tissues).

Abstract: Methods and compositions provide suitable support material for culturing cells with a desirable metabolic activity. For example, keratinocytes directly grown on flexible supports show metabolic activity. Wound healing methods and compositions using the cells grown on flexible supports, wherein the cells exhibit increased metabolic activity are disclosed.

Abstract: A tissue culture device includes a container having one or more upstanding walls extending upwardly from a floor. The floor of the container has a media having nutrients or growth substances therein. A plurality of plant tissues are within the container compartment and are placed upon a screen between the plant tissues and the media. The screen is tamped downwardly onto the media so that the plant tissues can get nutrients from the media, and so that waste products are transferred into the media. The screen is also removable through the open end of the container so as to remove all of the plurality of the plant tissues from the container at once. The plant tissue device can then be placed in another container having any cultured media as needed for the correct maintenance, propagation and development of plant tissue in culture.

Abstract: To produce a cell culture membrane having biocompatibility utilizing DNA of natural resources and a cell culture kit, the cell culture membrane having DNA ionically-cross-linked with calcium ions or magnesium ions is provided. To produce a porous material utilizing DNA, a production method for a cell culture membrane and a production method for a porous material, fine pores of 1 nm to 100 ?m in diameter are prepared in the porous material.

Abstract: The present invention relates to a method for preparing a cell-derived extracellular matrix membrane, more particularly, to a method for preparing a chondrocyte-derived ECM membrane, the method comprising the steps of forming a suitable thickness of ECM membrane by culturing chondrocytes derived from animal cartilage at a high concentration in vitro, and drying it after decellularization process. The cell-derived ECM membrane scaffold according to the present invention is composed of extracellular matrix secreted by chondrocytes so that the membrane has excellent biocompatibility as well as an immune-previlage effect specific to cartilage. Since the membrane also has a suitable compressive strength, it can be used to replace periosteum for cartilage regeneration or artificial collagen membrane and used as dura mater transplant material, a natural ECM membrane for treating skin loss, materials for cell transplantation and a growth factor delivery vehicle.

Abstract: The present invention relates to a method for fabricating a cell-derived extracellular matrix scaffold, more particularly, to a method for fabricating a cell-derived extracellular matrix scaffold, the method comprising the steps of obtaining a chondrocyte/extracellular matrix (ECM) membrane from chondrocytes derived from animal cartilage, obtaining a pellet-type scaffold-free construct by culturing after centrifuging the obtained chondrocytes/extracellular matrix (ECM) membrane and freeze-drying the obtained pellet-type construct. The cell-derived ECM scaffold according to the invention is a porous scaffold fabricated using cartilage tissue engineered by culturing chondrocytes in an in vitro scaffold-free system, which is not reduced in size during the cultivation and thus useful for cartilage regeneration.

Abstract: A disposable apparatus for cell expansion, having at least one bioreactor. The bioreactor has a cellular growth area and a supply area, the cellular growth area being separated from said supply area by a membrane. A fluid recirculation path in fluid communication with the cellular growth area allows for hermetically removing a sample containing cellular matter. This may comprise an elongated tube, or a plurality of parallel tube segments. The parallel tube segments have inflow ends and outflow ends, and the inflow ends are joined at a first common juncture and the outflow ends are joined at a second common juncture. The common junctures may comprise valves.

Abstract: A method for producing cell material (20) having multiple biological cells (21), which have a predefined geometrical arrangement, includes the steps of providing a manipulation tool (10) having a tool body (11), whose surface (12, 14) at least partially contacts the cell material (20), and adjusting the manipulation tool (10) using a change of geometrical properties of the surface (12, 14) in such a way that the geometrical arrangement of the cells (21) is changed. A manipulation tool for performing a method of this type is also described.

Abstract: The present invention provides a novel substrate for use in growing cells and for the study of mechanobiology. The membrane of the present invention comprises appropriate microtopography and surface chemical modifications to facilitate the production of adherent and oriented cells that phenotypically resemble cells in vivo.

Abstract: The present invention relates to methods of generating an ex vivo tissue-like system in a bioreactor system capable of supporting continuous production of, and output of cells and tissues and an ex vivo tissue system made therefrom.

Abstract: We describe a cell culture substrate comprising a polymerised high internal phase emulsion polymer adapted and modified for use in the routine culture of cells in three dimensions; typically mammalian cells and the use of the substrate in a cell culture system for investigation and analysis of proliferation, differentiation and function of cells.

Abstract: A method of pulsing cultured hepatocytes, such as sandwich-cultured hepatocytes. The method includes providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing the culture of hepatocytes to a calcium-free buffer, whereby the contents of the at least one bile canaliculus are released; and removing the calcium-free buffer Pulsing cultured hepatocytes can reduce cholestasis arid can provide an in vitro culture of hepatocytes the more closely reflects in vivo hepatocyte characteristics.

Abstract: A porous scaffold having pores for seeding cells characterized in that, in the outer peripheral face of the porous main body having the pores for seeding cells, a porous membrane having pores smaller than the cells is located. Thus, it is possible to provide a porous scaffold whereby the cells can be seeded at a high efficiency while preventing cell leakage and, moreover, even cells having little adhesiveness can be adhered.

Abstract: A nanoporous membrane structure characterised in that it has a hexagonal array of tubes with a substantially uniform inter-pore distance between the tubes wherein the distance is within the range from 10-500 nm and wherein the tubes may have a depth of up to 500 ?m.

Abstract: A method for supporting the in vitro growth of one or more eukaryotic cell type(s), the method comprising; seeding cells of said cell type(s) onto particles comprising basement membrane or a basement membrane-like substrate, and culturing said seeded cells in vitro under conditions suitable for expansion of said seeded cells, wherein said particles are less than about 500 ?m in size.

Abstract: The present invention provides a devitalized mammalian parenchymatous tissue composition which includes an interstitial structure which can serve as a scaffold for tissue repair or regeneration. The devitalized mammalian parenchymatous tissue composition can further include the basement membrane of the tissue.

Abstract: Provided are compositions and methods for in vitro generation and in vivo use of tissue for the repair of defective tissue, especially cartilage. Chondrocytes or other cells are cultured in vitro in a biodegradable amorphous carrier within the confines of a space bounded by a semi-permeable membrane with a molecular weight cut-off of greater than 100 kDa. The culture can be subjected to physical/physicochemical conditions that mimic in vivo conditions of the tissue in need of repair or replacement. In one embodiment the invention provides an amorphous preparation of chondrocytes and their extracellular products, suitable for injection.

Abstract: In general, the present invention is related to collagen compositions and thin films, and to methods of making and using the same. In some embodiments, the present invention is directed to “uniaxial pattern” or “linear pattern” collagen materials, compositions and thin films, and methods of making.

Abstract: The present invention relates to cell and tissue culture. In particular, the present invention provides a method for preparing an organotypic culture using dissociated cells or microexplants obtained from an animal organ. The method for preparing an organotypic culture comprises culturing cells from an organ on a surface characterised in that the cells are compacted. The invention further relates to a high-throughput method for the preparation of a collection of organotypic cultures. The invention further relates to a device for carrying out a method of organotypic culture according to the invention.

Abstract: A device for monitoring the migration or invasion of a biological particle such as a cell is disclosed. The device includes an upper chamber adapted to receive and retain a cell sample, a lower chamber having at least two electrodes, and a biocompatible porous membrane having a porosity sufficient to allow cells to migrate therethrough. The membrane is disposed in the device so as to separate the upper and lower chambers from one another. Migration of cells through the porous membrane permits contact between the migrating cells and one or more electrodes of the lower chamber. The contact provides a detectable change in impedance between or among the electrodes.

Abstract: Provided is a method for controlling the degree of labeling (DOL) of a carrier molecule or solid support by the addition of a reactive label competitor to the labeling reaction. When the reactive label competitor is added to the labeling solution the competitor competes with the carrier molecule or solid support for the label, reducing the number of labels available to conjugates to the carrier molecule or solid support. This provides for a facile method that predictably alters the DOL of a carrier molecule or solid support.

Abstract: A bio-artificial organ comprises a substrate comprising a roll of a substrate material, and a plurality of cells adhered to the substrate, the roll being formed from a sheet rolled to form a plurality of layers that include spacers and spaced openings such that at least a first set of parallel chambers is formed when the roll is formed, the chambers being manifolded to a first inlet and a first outlet. The bio-artificial organ may further include at least a second chamber, the second chamber being isolated from the first set of chambers by at least a cell barrier. A method for assembling a bio-artificial organ comprises a) providing a substrate for cell culture capable of forming a roll, the substrate having a surface, b) patterning the surface of the substrate, c) seeding cells onto the substrate, and d) reeling the substrate into a cylindrical roll.

Abstract: The present invention provides a three-dimensional tissue equivalent for in-vivo and in-vitro uses. The three dimensional tissue equivalent of the present invention is a non-contractile cellular sheet cultured over a porous scaffold using a macromass culturing technique, for example where the cellular sheet is entirely on one side of a porous sponge. In one embodiment, the present invention provides a dermal wound dressing that comprises a high density cellular sheet of dermal fibroblast cells.

Abstract: A method and apparatus for preparing biological cell samples for intracellular analysis. The invention is based upon the recognition that many of the steps of the conventional methods for such sample preparation can be eliminated, leading to a process that readily lends itself to automation and the advantages associated therewith. The method of the invention comprises the steps of (a) cell-fixation, (b) permeabilization and (c) staining (or labeling) of intracellular molecules of interest by probes that are readily detectable by flow cytometric techniques, all without any intervening cell-washing (and re-suspension) steps. Rather, the single cell-washing step is effected after these three steps have been carried out. Preferably, the washing step is carried out by passing the fixed, permeabilized and stained cell sample through a semi-permeable membrane that serves to filter out (by transmission) interferants to waste while retaining the cells of interest.

Abstract: The present invention provides a fusion protein which delivers a functional protein or peptide into a cell at enhanced efficiency. The fusion protein of the present invention is a transduction domain-target protein-transduction domain fusion protein, wherein the transduction domain, which comprises 6-12 amino acid residues whose more than ¾ consist of arginine or lysine residues, is covalently bonded to each of the amino- and carboxyl-terminal ends of the target protein. Green fluorescence protein and Cu/Zn-superoxide dismutate (SOD) are used as the target protein.

Abstract: A carrier for cell culture comprising a carrier having a cationic group and polypeptide-modified portions formed on a surface of the carrier in a sea-island structure, and a carrier for cell culture comprising a water-containing gel comprising alginic acid, wherein a surface of the carrier is coated with collagen, and wherein the collagen is bound to a surface of the water-containing gel by means of chitosan.

Abstract: Arrays including microparticles having probe and marker moieties are used for the detection of a target in a sample. Microparticles are randomly immobilized on at least a portion of a substrate. A detection scheme is performed to detect the marker associated with the microparticle and the identity of the probe, and any target bound to the probe.

Abstract: The invention relates to a reactor for cell and tissue culture particularly suited to the culture of cells or tissues in the form of thin layers, and comprises the means for ensuring a physiological stimulation and supply of nutrients in culture medium adapted to the reduced thickness of the culture. The reactor can be used for the preparation of an implant and also includes the means of ensuring a mechanical stimulation of tissues or cells and supplying nutrients in culture medium suitable for structural tissues. The reactor also comprises, in an advantageous way, the means or a form that allows easy introduction of the tissue or the solid part of an implant into the culture chamber and, for example, a wall or an elastically deformable element. In addition, the reactor can include means of temperature regulation. The reactor can be used for the culture of different types of cells and the preparation of implants in a variety of forms, compositions and applications.

Abstract: The present invention relates to methods for the transformation and regeneration of transformed embryogenic tissue of coniferous plants. In particular, the invention relates to improved methods for transforming embryogenic tissue of coniferous plants and for regenerating transformed embryogenic tissue of coniferous plants. The invention is well suited to the transformation and regeneration of transformed embryogenic tissue of plants of the subgenus Pinus of pines and hybrids thereof.

Abstract: Described herein is the self-assembly of amphiphilic peptides, i.e., peptides with alternating hydrophobic and hydrophilic residues, into macroscopic membranes. The membrane-forming peptides are greater than 12 amino acids in length, and preferably at least 16 amino acids, are complementary and are structurally compatible. Specifically, two peptides, (AEAEAKAK)2 (ARARADAD)2, were shown to self-assemble into macroscopic membranes. Conditions under which the peptides self-assemble into macroscopic membranes and methods for producing the membranes are also described. The macroscopic membranes have several interesting properties: they are stable in aqueous solution, serum, and ethanol, are highly resistant to heat, alkaline and acidic pH, chemical denaturants, and proteolytic digestion, and are non-cytotoxic.

Abstract: A carrier for cell culture comprising an alginic acid gel layer laminated with a gel layer containing a cell adhesion substance, wherein the gel layer containing a cell adhesion substance has a dry thickness of less than 0.3 ?m. The carrier for cell culture enables reproducible and convenient cell layer lamination without inhibiting growth and proliferation of cells upon solubilization of the carrier for preparation of a cell sheet.

Abstract: A carrier for cell culture comprising a water-containing polymer gel containing chitosan, wherein the water-containing polymer gel is coated with collagen and/or alginic acid, and a carrier for cell culture, which comprises a gel layer containing chitosan and an inorganic layer adjacently provided to the gel layer.

Abstract: An apparatus (10) to grow cell cultures includes an anchor (14) potted to a flexible membrane (12) and an anchor stem (16) to which cells attach. A jig (18) having a trough (46) and at least one passage (52) is positioned adjacent the flexable membrane (12). A method for growing and mechanically conditioning three-dimensional cell constructs begins by drawing the flexible membrane (12) and the anchor stem (14) into the trough (46) by means (20) for flexing the flexible membrane (12) (step 114). Cells are supplied (step 116) and allowed to attach to the anchor stem (16) (step 118). The flexible membrane (12) is released from within the trough (46) (step 120), resulting in a three-dimensional structure (34) of cells attached to the anchor stem (16). The cells grow into the three-dimensional construct (step 130). The structure (34) and/or the construct can be subjected to a regimen of strain for mechanical conditioning of the cells (step 132).

Abstract: The invention relates to a novel method, permitting the semiquantitative determination of the confluence of a cell layer on the total surface of a porous biomaterial, without irreversible damage to part or indeed all of the hybrid construct. A method is described for the first time, which is able to detect the formation of the confluence or the confluence itself, without destroying the hybrid construct, and thus permits the finished hybrid product to be withdrawn from the essential growth conditions only after the determination of the completed confluence, and to immediately direct it to its further use.

Abstract: The present invention provides a novel substrate for use in growing cells and for the study of mechanobiology. The membrane of the present invention comprises appropriate microtopography and surface chemical modifications to facilitate the production of adherent and oriented cells that phenotypically resemble cells in vivo.

Abstract: Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

Abstract: A method for obtaining lineage committed human cells imbued with enhanced proliferative potential, biological function, or both, comprising culturing lineage committed human cells under physiologically acceptable liquid culture conditions, where the liquid culture medium is replaced at a rate and for a time sufficient to obtain the human lineage committed cells imbued with enhanced proliferative potential, biological function, or both; and isolating the cultured cells.

Abstract: An apparatus and method for the reconstruction of a previously torn ligament using a tissue-engineered ligament. The tissue-engineered ligament includes a scaffold of biocompatible material having at least one layer and forming a sheet. The scaffold is placed in a cultured medium for seeding with fibrocyte forming cells. The seeded scaffold is then placed in an incubator to increase the number of cells. The seeded scaffold is then formed into a slender structure suitable for implantation. The method of making a tissue-engineered ligament includes forming a scaffold of biocompatible material having at least one layer forming a sheet. Next, the scaffold sheet is seeded with fibrocyte forming cells. The method further includes increasing the number of cells on the seeded scaffold and forming a slender structure suitable for implantation from the scaffold.

Abstract: Multiple cell layers are formed with one cell layer formed on another cell layer. A carrier is provided having an alginate gel layer formed on a porous membrane. An extracellular matrix component gel layer or extracellular matrix component sponge layer may be formed on the alginate gel layer. A cell layer is formed on the alginate gel layer, or the extracellular matrix component gel layer or extracellular matrix component sponge layer. The alginate gel layer is solubilized such as with a chelating agent to exfoliate the cell layer from the porous membrane, and the exfoliated cell layer is placed on another cell layer on a carrier. The number of cell layers formed on each other may be 1-10, preferably 1-5, and more preferably 1-3.

Abstract: A method and structure for growing living organic tissue in which a scaffold is provided on which cells of the tissue may be seeded and on which the tissue may be grown and a plurality of membrane capillaries are provided for conveying a nutrient fluid, the plurality of membrane capillaries being interspersed through the scaffold and having membranes which are permeable to nutrients and oxygen such that the tissue may be grown throughout the scaffold and whereby upon in vivo implantation a supporting nutrient tissue supply rapidly establishes.

Abstract: Described herein is the self-assembly of amphiphilic peptides, i.e., peptides with alternating hydrophobic and hydrophilic residues, into macroscopic membranes. The membrane-forming peptides are greater than 12 amino acids in length, and preferably at least 16 amino acids, are complementary and are structurally compatible. Specifically, two peptides, (AEAEAKAK)2 (ARARADAD)2, were shown to self-assemble into macroscopic membranes. Conditions under which the peptides self-assemble into macroscopic membranes and methods for producing the membranes are also described. The macroscopic membranes have several interesting properties: they are stable in aqueous solution, serum, and ethanol, are highly resistant to heat, alkaline and acidic pH, chemical denaturants, and proteolytic digestion, and are non-cytotoxic.

Abstract: A method and apparatus for screening an array of test compounds for bioactivity by contacting an array of test compounds with a detector layer capable of detecting bioactivity, and detecting a detector layer response. The detector layer is comprised of physiologically viable cells. The method and apparatus allow a large number of test compounds to be simultaneously assayed in parallel without the need for complex fluidic devices.

Abstract: Rolling circle amplification (RCA) has been useful for detecting point mutations in isolated nucleic acids, but its application in cytological preparations has been problematic. By pretreating cells with a combination of restriction enzymes and exonucleases, we demonstrate RCA in solution and in situ to detect gene copy number and single base mutations. It can also detect and quantify transcribed RNA in individual cells, making it a versatile tool for cell-based assays.

Abstract: The invention features modular cell culturing devices including one or more flat-plate modules, and is based on the discovery that if the flows of liquid medium and oxygenated fluid are separated by a gas-permeable, liquid-impermeable membrane, and the cells are grown attached to the liquid side of the membrane, the device can be used to culture cells with transport of oxygen through the membrane (i.e., direct oxygenation), without regard for the flow rate of the liquid medium passing through the device. The new flow-through cell culturing devices can thus be used to culture cells, e.g., hepatocytes, with high levels of cell function in organ, e.g., liver, assist systems, for production of cells, for production of cell-derived products, such as, proteins or viruses, or for systems to treat biological liquids to remove toxins, such as, ammonia, or add cell-synthesized products, or both.

Abstract: An apparatus and method for purification and assay of neurites is useful for separation and analyses of extension organelles and/or protrusion of cells for purification, production, observation, and quantification of neurites in the neurobiology field. The present invention provides a pore-sized controlled porous filter membrane which outspace side surface is coated with a cell adhesion layer to form an adhesion surface, and combines the neuronal cells with the porous filter membrane in an aqueous environment under conditions in which outgrown neurites from cell bodies of the neuronal cells are attracted to and grow on the adhesion surface, wherein the outgrown neurites of the neuronal cells pass through pores provided in the porous filter membrane to the adhesion surface while each of the pores has a size smaller than the cell bodies of the neuronal cells so as to prevent the cell bodies of the neuronal cells passing through the pores and remaining on an opposing side surface of the porous filter membrane.

Abstract: Methods and apparatus are provided for forming a vascular graft in vitro by axially distending a blood vessel to induce growth. The apparatus comprises a chamber containing a tissue culture medium, an inlet cannula, an outlet cannula, and a means for moving the inlet cannula, the outlet cannula, or both, to axially stretch a donor blood vessel secured between the inlet cannula and the outlet cannula in a submerged position in the tissue culture medium, wherein the inlet cannula, the outlet cannula, and the donor blood vessel are secured together to form a conduit through which the tissue culture medium can flow.