Abstract: The present invention relates to cell adhesion promoting (“CAP”) peptide combinations that promote cell attachment or cell adhesion to culture surfaces that are otherwise cell adhesion resistant “CAR”. The invention provides combination of peptides that, when covalently coupled to a CAR layer such as hyaluronic acid that has been created on a polystyrene surface, promote cell attachment, growth differentiation, and execution of other desired cellular functions in culture.

Abstract: The invention discloses methods of proliferation and differentiation of multipotent neural stem cells. Also provided are methods of making cDNA libraries and methods of screening biological agents which affect proliferation differentiation survival phenotype or function of CNS cells.

Abstract: A process for producing a film for controlling the chemotactic function in an extremely small area in which a chemotactic factor substance in the area has a concentration gradient in one direction. The invention further includes an artificial material with an arrangement in which, on a substrate, there is a film having a chemotactic factor substance for controlling the chemotactic function which it has a concentration gradient in one direction. The process of the invention includes controlling the amount of irradiation in one direction of the film such that the chemotactic factor substance is degenerated which results in a corresponding concentration gradient of the film in the one direction.

Abstract: This invention relates to the discovery that an intermediate, differentiated stage of pancreatic stem cells exist that can be matured in situ into a stable cell line that produces insulin in response to glucose. These cells are advantageous in that they are both expandable and stable in culture. This invention avoids the step of culturing the intermediate stage stem cells into later stage pancreatic cells.

Abstract: Described is a deliverable composition with low toxicity comprising an amphipathic compound, a polycation, and a siRNA. The composition may be used in the process of delivering a siRNA to an animal cell or more particularly, a mammal cell.

Abstract: The present invention relates to cell adhesion promoting (“CAP”) peptide combinations that promote cell attachment or cell adhesion to culture surfaces that are otherwise cell adhesion resistant “CAR”. The invention provides combination of peptides that, when covalently coupled to a CAR layer such as hyaluronic acid that has been created on a polystyrene surface, promote cell attachment, growth differentiation, and execution of other desired cellular functions in culture.

Abstract: A novel cell culture medium suitable for primary culture of insect cells, an insect-derived water-soluble chitin, and a process of preparing an insect culture cell line in a short period of time by using the insect primary culture medium and the insect-derived water-soluble chitin. The insect cell primary culture medium comprises lactalbumin hydrolysate, yeastolate, and tryptose phosphate broth as protein extracts, and polyvinylpyrrolidone as a viscosity-supplementing agent. The insect-derived water-soluble chitin is subjected to deacetylation as the sole chemical modification.

Abstract: The invention provides a device for adhering cells in a specific and predetermined position, and associated methods. The device includes a plate defining a surface and a plurality of cytophilic islands that adhere cells, isolated by cytophobic regions to which cells do not adhere, contiguous with the cytophilic islands. The islands or the regions or both may be formed of a self-assembled monolayer (SAM).

Abstract: The invention pertains to methods and devices for the long term, in vitroculture of hematopoietic progenitor cells in the absence of exogenously added hematopoietic growth factors, improved methods for the introduction of foreign genetic material into cells of hematopoietic origin, and to apparatus for performing these methods. The hematopoietic progenitor cells are cultured on a three-dimensional porous biomaterial. The three-dimensional porous biomaterial enhances hematopoietic progenitor cell survival and leads to an expansion of progenitor cell numbers and/or functionality, while maintaining progenitor cell pluripotency in the absence of exogenous growth factors. In addition, the three-dimensional porous biomaterial supports high level transduction on cells cultured upon such environment.

Abstract: The invention concerns peptide fragments of the WASP family proteins, or peptides derived from said fragments, and their uses in particular for preparing reagents for use in implementing a method for detecting or screening molecules with inhibiting or stimulating effect on the formation of the actin cytoskeleton, hence an inhibiting or stimulating effect on cellular motility.

Abstract: A nucleic acid-bound polypeptide produced by binding a nucleic acid to a polypeptide, a method of producing the nucleic acid-bound polypeptide, and applications of the nucleic acid-bound polypeptide, including immunoassays for an antigen or antibody, such as an agglutination immunoassay are provided.

Abstract: The present invention provides specific peptides identified as having cell adhesion, growth, expression or secretion-enhancing activities. Many of the peptides of the invention may be produced in large quantity by such means as chemical synthesis or recombinant DNA methodology. They may be non-specifically adsorbed, or chemically attached to a surface or, alternatively, formulated in a culture medium to produce the desired effect on cultured cells.

Abstract: A polymer scaffold is provided comprising an extensively interconnected macroporous network. The polymer scaffold embodies macropores having a diameter in a range of 0.5-3.5 mm, and preferably in a range of about 1.0-2.0 mm. The polymer scaffold is prepared using a novel process which advantageously combines the techniques of particulate leaching and phase inversion to render a process that provides amplified means by which to control the morphology of the resulting polymer scaffold. The polymer scaffold has utility in the area of tissue engineering, particularly as a scaffold for both in vitro and in vivo cell growth. The polymer scaffold may be produced using pure polymer or alternatively a composite material may be formed consisting of a macroporous polymer scaffold and osteoclast-resorbable calcium phosphate particles with a binding agent binding the calcium phosphate particles to the polymer scaffold.

Abstract: A method and an apparatus for preparing and culturing cells, particularly bone marrow stromal cells. The method includes the steps of placing an oxygenator and a scaffold in a container, such as a syringe, withdrawing bone marrow stromal cells with the syringe, evenly distributing the cells on the scaffold, connecting the syringe with a reservoir with a medium to enrich the syringe with the medium, and promoting the medium level movement in the syringe to culture bone marrow stromal cells.

Abstract: Described are compositions with tethered growth effector molecules, and methods of using these compositions for growing cells and tissues. Growth effector molecules, including growth factors and extracellular matrix molecules, are flexibly tethered to a solid substrate. The compositions can be used either in vitro or in vivo to grow cells and tissues. By tethering the growth factors, they will not diffuse away from the desired location. By making the attachment flexible, the growth effector molecules can more naturally bind to cell surface receptors. A significant feature of these compositions and methods is that they enhance the biological response to the growth factors. The method also offers other advantages over the traditional methods, in which growth factors are delivered in soluble form: (1) the growth factor is localized to a desired target cell population; (2) significantly less growth factor is needed to exert a biologic response.

Abstract: The invention is directed to methods of making bioremodelable graft prostheses prepared from cleaned tissue material derived from animal sources. The bioengineered graft prostheses of the invention are prepared using methods that preserve cell compatibility, strength, and bioremodelability of the processed tissue matrix. The bioengineered graft prostheses are used for implantation, repair, or use in a mammalian host.

Abstract: The invention relates to a novel compound termed NKp30 that is selectively expressed by all mature NK cells and that is involved in human natural cytoxicity as an activatory receptor, to new antibodies that bind to the NKp30 structure, and to the pharmaceutical and medicinal uses thereof.

Abstract: A method for rendering a surface covered by a plastic material more hydrophilic by treatment with a gas plasma non-polymerizable gas. The method is characterized in that the intensity of the plasma is selected so that the surface becomes permanently more hydrophilic. Also disclosed is a naked plasma treated surface of plastic having an immediate water contact angle of ?30°, said water contact angle is changed ±20 and/or less than 5% upon washing with an ethanol/water mixture (70% w/w).

Abstract: The present invention relates generally to growth factors and more particularly to growth factors which are capable of stimulating or otherwise facilitating formation of insulin-secreting cells. The identification of these growth factors permits the development of protocols to culture cells in vitro for transplantation into mammalian and in particular human subjects with insulin-dependent type 1 diabetes or related conditions. It is further contemplated that the endogenous expression of growth factors required for the development of insulin-producing cells may be manipulated in vivo, by the appropriate administration of agents including genetic agents capable of regulating the expression of growth factors in pancreatic duct epithelial cells. The growth factors ray also be administered to subjects with type 1 diabetes to stimulate the proliferation and differentiation of pancreatic cells into insulin-secreting cells.

Abstract: A chimeric skin comprising immortalized human keratinocyte cells cocultured with donor keratinocytes is disclosed.

Abstract: New methods for producing tissue engineered constructs and engineered native tissues are disclosed. The methods include producing a tissue engineered construct by growing cells in vitro on a substrate and then decellularizing the construct to produce a decellularized construct consisting largely of extracellular matrix components. The construct can be used immediately or stored until needed. The decellularized construct can be used for further tissue engineering, which may include seeding the construct with cells obtained from the intended recipient of the construct. During any of the growth phases required for production of the construct, the developing construct may be subjected to various tissue engineering steps such as application of mechanical stimuli including pulsatile forces. The methods also include producing an engineered native tissue by harvesting tissue from an animal or human, performing one or more tissue engineering steps on the tissue, and subjecting the tissue to decellularization.

Abstract: The invention concerns a process for producing a three-dimensional bioartificial tissue having viable cells in or on a matrix, and by which cells and matrix can be cultivated into a tissue or a precursor of a tissue, a vascularized tissue of biological materials, obtained by this process, and an experimental reactor for scientific purposes and for producing clinically usable tissues and organs.

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: Implantable medical devices that include a non-woven framework are described, as well as methods of using such devices to deliver therapeutic compounds to a patient.

Abstract: The present invention pertains to a method for determining the sequence of a polynucleotide, the method relying on the detection of a conformational change in an enzyme that interacts with and processes along the polynucleotide. The detection of a conformational change may be carried out by measuring changes in a fluorophore bound to the enzyme.

Abstract: A polymer scaffold is provided comprising an extensively interconnected macroporous network. The polymer scaffold embodies macropores having a diameter in a range of 0.5-3.5 mm, and preferably in a range of about 1.0-2.0 mm. The polymer scaffold is prepared using a novel process which advantageously combines the techniques of particulate leaching and phase inversion to render a process that provides amplified means by which to control the morphology of the resulting polymer scaffold. The polymer scaffold has utility in the area of tissue engineering, particularly as a scaffold for both in vitro and in vivo cell growth.

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: Artificial liver devices and methods for using the devices to purify a biological fluid are disclosed. The methods include the use of living hepatocytes (23) which are either unattached or attached to inert carriers and suspended in a cell culture medium which circulates in the devices with the hepatocytes (23). Blood or plasma passes on one side (7?) of semi-permeable membranes, on the other side (7) of which is the cell culture medium and across which is a concentration and/or pressure gradient. Solutes diffusing across the membrane into the cell culture medium are metabolized by the hepatocytes (23) and/or captured by additional removal means (4). Those undesirable substances which do not diffuse out of the blood or plasma into the hepatocyte containing culture medium are captured by additional removal means (50).

Abstract: A process is provided for expanding the population of endothelial cells obtained from peripheral blood which can be transformed with a vector comprising a DNA sequence encoding a preselected bioactive polypeptide. The resulting transgenic endothelial cells are useful to biocompatibilize implantable medical devices or can be used directly, as for gene therapy.

Abstract: The invention relates to a bioreactor as well as a method for cultivating organic material: in particular cells, by means of a nutrient medium. For an intensive cultivation of the organic material in a simple and reliable handling a flow generating device is provided in the bioreactor according to the invention, by means of which the nutrient medium can be put into a flow, wherein a receiving device is arranged in the flow that is adapted for receiving and/or retaining the organic material and wherein the receiving device that is adapted for passing through the flowing nutrient medium is permeable. The method in accordance with the invention is characterized in that the nutrient medium is at least temporarily put into a flow, that the organic material, in particular cells, is retained in or at a receiving device, which is permeable to the nutrient medium and that the nutrient medium is passed through the receiving device.

Abstract: A spontaneously immortalized human keratinocyte cell line is disclosed. In a preferred embodiment, this cell line is ATCC 12191. In another embodiment of the invention, a method of assaying the effect of a test tumor cell modulation agent is disclosed. The method comprises the steps of obtaining a human stratified squamous epithelial cell culture, wherein the culture comprises human malignant squamous epithelial cells and spontaneously immortalized human keratinocytes, wherein the culture forms a reconstituted epidermis. One then treats the epidermis with a test tumor cell modulation agent and evaluates the growth of the malignant cells within the epidermis.

Abstract: The main object of the present invention is to provide a cell culture base material having a base material and a pattern of regions of excellent cell adhesiveness formed on the base material with high precision, and a method of producing the same.

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: The object of the present invention is to elucidate a biologically active function of a component constituting undegraded sericin and to provide a novel medical material, cosmetic material, etc. utilizing the functional composition. Disclosed is a cell growth promoter obtainable by elution from a fiber discharged by a domestic silkworm, e.g., cocoon filaments or the like, wherein the cell growth promoter comprises sericin having a molecular weight of about 400,000 as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a main component. This cell growth promoting agent (substance) is extremely useful because growth of cells is promoted when it is used in a wound dressing material, a vascular endothelium forming material and an organ forming material for medical use, in a cell culture base material for biological use, or in a cosmetic material for skin care use.

Abstract: Autologous cultured keratinocytes are grown on a biosynthetic and biocompatible substratum following pre-seeding with autologous or allogenic dermal fibroblasts. The resultant composite material may then be applied on the neodermis of artificial skin which had been previously engrafted on the patient. The composite material, and specifically Composite Biocompatible Skin Graft (CBSG) material comprises autologous keratinocytes and allogenic or autologous dermal fibroblasts grown on Laserskin. A method for cultivating the CBSG includes the application of dermal fibroblasts onto the substratum as a feeder layer and then the inoculation of autologous keratinocytes on the resultant structure. A method for engraftment comprises first applying an artificial skin with a protective silicone membrane on a wound area, thereby allowing vascularization; and following vascularization, removing the silicone membrane and engrafting the CBSG material onto the vascularized artificial skin.

Abstract: A mammalian muscle construct and a method for producing the construct are provided. The mammalian muscle construct includes a substrate and a plurality of separate anchors secured to the substrate. Myogenic precursor cells are provided on the substrate with at least some of the cells in contact with the anchors. The myogenic precursor cells are cultured in vitro under conditions to allow the cells to become confluent between the anchors. The anchors are receptive to the cells and allow the cells to attach thereto, such that placement of the anchors controls the size and shape of the muscle construct formed. Specifically, the anchors include separate fragments of biocompatible material secured to the substrate, wherein cell adhesion molecules are associated with each fragment to facilitate attachment of the precursor cells to the fragment.

Abstract: This invention relates to the discovery that an intermediate, differentiated stage of pancreatic stem cells exist that can be propagated in a stable manner in successive serial passaging while maintaining insulin production in response to glucose. These cells are advantageous in that they are both expandable and stable in culture and can driven to late stage development, i.e. prototype islet cells. This invention further provides for culturing techniques that select for these intermediate differentiated stage cells and selectively eliminates early or late stage pancreatic cells.

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: The present invention relates generally to tissue implant material for use in grafting procedures. More particularly, the present invention provides non-vascular tissue for use as vascular graft material. The present invention further contemplates a method of vascular grafting using non-vascular tissue. The tissue of the present invention is preferably autologous relative to the recipient of the graft and is conveniently prepared around or on a molding support or other foreign body inserted into a body cavity of the intended recipient of the graft. The tissues and methods of the present invention are particularly useful in the treatment or prophylaxis of diseased or damaged blood vessels such as in atherosclerosis.

Abstract: A method of preparing a protein array based on biochemical protein-protein interaction is provided. An array of a first protein which includes a PDZ domain is deposited on a substrate. A second protein, which includes an amino acid sequence (S/T)—X—(V/I/L)—COOH (each hyphen represents a peptide bond, each parenthesis encloses amino acids which are alternatives to one other, each slash within such parentheses separates the alternative amino acids, and the X represents any amino acid which is selected from the group comprising the twenty naturally occurring amino acids), is applied to the first protein array. The amino acid sequence (S/T)—X—(V/I/L)—COOH of the second protein is bound to the PDZ domain of the first protein.

Abstract: The invention relates to substrates containing polyphosphazene with a forming surface as matrices for producing biological materials that can be implanted in a mammal. The invention also relates to a method for producing said substrates and substrates containing polyphosphazene with a microstructured surface.

Abstract: A method is provided for long term culture of proliferating hepatocytes that retain hepatic function to produce a hepatic cell culture. Hepatocytes and nonparenchymal cells are co-cultured ex vivo on a matrix coated with a molecule that promotes cell adhesion, proliferation or survival, in the presence of growth factors, resulting in a long-term culture of proliferating hepatocytes that retain hepatic function. The co-culturing method results in the formation of matrix/hepatic cell clusters that may be mixed with a second structured or scaffold matrix that provides a three-dimensional structural support to form structures analogous to liver tissue counterparts. The method can be used to form bio-articial livers through which a subjects blood is perfused.

Abstract: A method for adhering and proliferating cell, which comprises the steps of inoculating, culturing and then killing fibroblast derived from a mammal, is provided. A culture vessel manufactured according to the steps of the method which can provide improved adhesion to cell and enhanced cell-proliferation is also provided.

Abstract: Methods and compositions are provided for the treatment of articular cartilage defects and disease involving the combination of tissue, such as osteochondral grafts, with active growth factor. The active growth factor is preferably a composition containing at least one bone morphogenetic protein and a suitable carrier. The method results in the regeneration of functional repair of articular cartilage tissue.

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: The present invention relates to a reactor module constructed of hollow fibers and cells as well as to reactors comprising this reactor module.

Abstract: Various embodiments of the present invention provide both apparatus and method for the patterning of cells onto non-adhesive substrates and the controlled regulation of cells through the use of photoactivated cellular morphogenic factors.

Abstract: A cell and substrate system and nerve regeneration implant are disclosed including a carbon nanotube and a neuron growing on the carbon nanotube. Both unfunctionalized carbon nanotubes and carbon nanotubes functionalized with a neuronal growth promoting agent may be utilized in the invention. A method is also disclosed for promoting neuronal growth.

Abstract: The present invention is directed, in certain embodiments, to improved, small scale systems and methods able to selectively treat parts of a single cell, including, in certain embodiments, portions of a main body portion of a single cell, and able, in certain embodiments, to establish long-term gradients of active substances within subcellular regions of a single cell. The present invention provides, in some embodiments, techniques for selectively contacting a portion of the surface of a biological cell with a fluid or fluid component carrying a particular potential for a biophysical or biochemical interaction with the cell, and simultaneously contacting a different portion of the surface of the cell with another fluid or fluid component having a different potential for the biophysical or biochemical interaction with the cell.

Abstract: The invention pertains to methods and devices for the long term, in vitroculture of hematopoietic progenitor cells in the absence of exogenously added hematopoietic growth factors, improved methods for the introduction of foreign genetic material into cells of hematopoietic origin, and to apparatus for performing these methods. The hematopoietic progenitor cells are cultured on a three-dimensional porous biomaterial. The three-dimensional porous biomaterial enhances hematopoietic progenitor cell survival and leads to an expansion of progenitor cell numbers and/or functionality, while maintaining progenitor cell pluripotency in the absence of exogenous growth factors. In addition, the three-dimensional porous biomaterial supports high level transduction on cells cultured upon such environment.