Abstract: The present invention provides cultured leukemia cells. The method comprises isolating mononuclear cells, which contain leukemia cells, culturing the leukemia cells in a chamber having a scaffolding covered or surrounded with culture medium, where the scaffolding allows for leukemia cells to have cell to cell contacts in three dimensions. The subject leukemia cells are useful for screening compounds which inhibit or stimulate leukemia cell function or formation.

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 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: A bioreactor for cell culture includes a chamber having three channels therethrough for conducting fluids, the three channels including an upper channel, a middle channel, and a lower channel, each the channel having an inlet and an outlet approximately opposite along the channel from the inlet. First and second cell support scaffolds are positioned within the chamber, each scaffold comprising at least one three-dimensional porous matrix containing non woven fibrous polyethylene terephthalate, the first scaffold being positioned within the chamber between the upper channel and the middle channel, and the second scaffold being positioned within the chamber between the middle channel and the lower channel.

Abstract: An apparatus and method for a modular cell culture bioreactor comprises a plurality of chambers for cell culture; at least one reservoir containing a cell support medium; a plurality of conduits fluidly connecting the at least one reservoir with the plurality of chambers; and at least one pump fluidly connected through the plurality of conduits with the at least one reservoir and with the plurality of chambers to pump cell support medium therethrough; wherein each individual chamber of the plurality of chambers includes at least one three-dimensional matrix comprising polyethylene terephthalate, a plurality of channels carrying the cell support medium and having the matrix positioned in fluid communication therebetween, and at least two openings into each the channel, wherein a first the opening is in fluid connection with the pump and a second the opening is in fluid connection with the reservoir.

Abstract: In a system for generating tissue by growing cells in a porous and sometimes biodegradable material, a needle punched textile which serves as a scaffold is used for growing any of a variety of anatomical elements, in which the thickness of areas of the anatomical element and thus its strength can be increased by providing layers of mesh which are needled together to form a layerless textile and in which delamination is prevented through the use of the needling. In one embodiment, the needle punched textile is utilized to form a semi-lunar heart valve. In a preferred embodiment for pediatric use, the textile is made from two different biodegradable non-woven meshes. For some adult applications biodegradable meshes are not necessary, thus eliminating the necessity of using two different needled meshes. In one embodiment the needling is done with a single needle which is made to move around the periphery of a mold used in making the scaffold, thus to precisely control the area needled.

Abstract: The present invention provides nano-porous fibers and protein membrane compositions. In certain embodiments, continuous fiber compositions are provided having nanometer sized diameters and surface pores. In another embodiment, a protein membrane composition is provided comprising a protein; and a polymer, wherein the protein and the polymer are electrospun to form a protein membrane composition. In certain instance, the protein is covalently bound to the fiber.

Abstract: A composite scaffold for engineering a heterogeneous tissue is provided. The composite scaffold includes: (a) a first scaffold being capable of supporting, formation of a first tissue type thereupon; and (b) a second scaffold being capable of supporting formation of a second tissue type thereupon; wherein the first scaffold and the second scaffold are arranged with respect to each other such that when the first scaffold supports the first tissue type and the second scaffold supports the second tissue type, a distance between any cell of the second tissue type and the first tissue type does not exceed 200 $G(m)m.

Abstract: A bioreactor for three-dimensional culture of liver cells is disclosed. The device is characterized by the use of textile vasculatures. A model and method for optimizing vasculature parameters is also disclosed. Liver acinar structure and physiological parameters are mimicked by sandwiching cells in the space between the two innermost woven textile hollow fibers, and creating radial flow of media from an outer compartment, through the cell mass compartment, and to an inner compartment. The theoretical optimum hydraulic permeability for the two innermost semi-permeable membranes is determined based on physiological hepatic sinusoidal blood flow and pressures. Experimental studies using a flow rate and pressure monitoring systems in conjunction with phase-contrast velocity-encoded MRI confirm theoretical results. Novel woven vascular tubes with optimum hydraulic permeability are disclosed for culturing hepatocytes in the multi-coaxial bioreactor.

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: A relic process is used to produce bioresorbable ceramic scaffolds that can be used for in vitro or in vivo growth of human or animal tissue such as bone or cartilage. The process involves impregnating an organic fabric template with metal and phosphate ceramic precursors, heat treating the impregnated fabric to decompose the fabric to form a ceramic green body, and sintering the ceramic green body to form the scaffold which has a form analogous to that of the fabric template. Impregnating the fabric may be by soaking the fabric in a solution or sol containing the ceramic precursors. The fabric may be formed into a laminate prior to heat treating. Sintering results in fibers of the fabric being cross-sintered with one another to form a three-dimensional scaffold structure having controlled pore size and distribution. The scaffold may be treated with a material that promotes bone growth through the scaffold.

Abstract: A method of preparing a placental-derived amniotic membrane biofabric is provided. The biofabric is a dry decellularized amniotic membrane that is capable of being stored at room temperature, and subsequent to rehydration can be used for a variety of medical and/or research purposes. A laminate of said biofabric is also provided that can be shaped into complex shapes and repopulated with cells to generate both acellular and cellularized engineered tissues and organoids.

Abstract: The present invention provides a method for preparing biodegradable porous polymer scaffolds for tissue engineering, comprising: a) fabricating a polymer sample from a polymer solution containing at least one biodegradable polymer and an effervescent mixture; b) effervescing the polymer sample in the presence of an effervescent medium such as an aqueous alcohol solution; and c) drying. The method for preparing biodegradable polymer scaffolds of the present invention has the advantages that the process is simple, that pore size can be easily controlled, that the problem caused by the secretion and existence of the toxic substance can be avoid by using a material harmless to human body, and that high efficiency can be achieved. In addition, biodegradable porous polymer scaffolds prepared by above method have the advantages that high porosity can be achieved and an open cell structure in which pores are interconnected is obtained.

Abstract: A composite material is provided that exhibits excellent biocompatibility, is easy to handle in clinical applications, and has an excellent mechanical strength. The composite material can be used as a scaffold for supporting chondrocytes or progenitor cells differentiating thereto and is useful for an implant for cartilage tissue regeneration.

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: Methods and compositions for a native-state histoculturing system for skin are disclosed. Skin samples are placed on an extracellular support matrix immersed in a medium whereby the internal surface of the skin is adjacent to the matrix and the external surface is exposed above the surface of the medium and the skin maintained in the medium under skin culturing conditions. An extracellular support matrix comprising a collagen-containing gel and homopolysaccharide sponge or a combination matrix of a top layer of a collagen-containing gel and a bottom layer of a homopolysaccharide sponge is also disclosed. Skin toxicity, hair growth, anti-aging and anti-wrinkling assays utilizing the histoculturing system of the present invention are also disclosed.

Abstract: The present invention discloses a carrier for cell attachment or fixation and its preparation. The carrier of the present invention is formed by the following steps: forming a fiber by extruding a melted polymer from a nozzle: extending the fiber and shaping the extended fiber on the shaping net to form a three-dimensional branch-like non-woven structure; activating the surface of the non-woven structure to have cell affinity and facilitate the attaching and growing of the cell. Moreover, the surface of the carrier is wrinkled or rough to improve the stiffness of the carrier and keep enough space between the carriers.

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 patent describes a three-dimensional interconnected open cell porous foams that have a gradient in composition and/or microstructure through one or more directions. These foams can be made from a blend of absorbable and biocompatible polymers that are formed into foams having a compositional gradient transitioning from predominately one polymeric material to predominately a second polymeric material. These gradient foams are particularly well suited to tissue engineering applications and can be designed to mimic tissue transition or interface zones.

Abstract: Apparatus and methods are disclosed for maturing an elongate replacement tissue construct in vitro prior to use of the replacement construct in vivo as, for example, a ligament. The tissue is seeded with specific cells, exposed to a maturation fluid, and subjected to selected forces, which can include longitudinal stress, (i.e. stressing the tissue along its elongate axis). The tissue is disposed in a maturation chamber that confines maturation fluid for introduction to the tissue. A first mounting element couples to a first end of the elongate biopolymer tissue and a second mounting element couples to a second end of the tissue such that the tissue extends along a longitudinal axis, and a force is applied to at least one of the mounting elements for longitudinally stressing the tissue. The foregoing apparatus and methods are intended to provide a replacement tissue that is more readily integrable in vivo, i.e.

Abstract: Cartilage tissue and implants comprising tissue are produced in vitro starting from cells having the ability to form an extracellular cartilage matrix. Such cells are brought into a cell space (1) and are left in this cell space for producing an extracellular cartilage matrix. The cells are brought into the cell space to have a cell density of ca. 5×107 to 109 cells per cm3 of cell space. The cell space (1) is at least partly separated from a culture medium space (2) surrounding the cell space by means of a semi-permeable wall (3) or by an open-pore wall acting as convection barrier. The open-pore wall can be designed as a plate (7) made of a bone substitute material and constituting the bottom of the cell space (1). The cells settle on such a plate (7) and the cartilage tissue growing in the cell space (1).

Abstract: Methods and compositions are described that provide three-dimensional fibrillar matrices useful as, among other things, structural prosthetics and scaffolds for cells. The porous fibrillar matrices of the present invention have desirable mechanical properties suitable to a variety of applications, including platforms for in vitro cell cultivation, implants for tissue and organ engineering, implants as tendon and facia prosthetics, and product packaging.

Abstract: A stromal cell-based three-dimensional cell culture system is provided which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. The stromal cells along with connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope a framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. Living stromal tissue so formed provides support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and/or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo, which can be utilized in the body as a corrective tissue.

Abstract: Compositions and methods are provided for adhering and binding biologically active proteins and protein-containing composites to substrates. Adhesive formulations comprising a nonproteinaceous polymer of monomeric units comprising an aromatic moiety substituted with at least one hydroxyl group such as poly(p-hydroxy-styrene) are applied to substrates and subsequently contacted with proteins. Beads comprising a nonproteinaceous polymer of monomeric units comprising an aromatic moiety substituted with at least one hydroxyl group are also provided, and the beads are coated with a protein. Substrates to which the adhesive formulations have been applied, as well as the beads, can be used to adhere cells and tissues, to sort cell types, to perform immunoassays, to perform chromatography and to remove protein from samples.

Abstract: There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.

Abstract: Disclosed 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 new 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: A stromal cell-based three-dimensional cell culture system is prepared which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. The stromal cells and connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope a framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. The living stromal tissue so formed provides the support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and/or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo, which can be utilized in the body as a corrective tissue.

Abstract: A perfusion device such as a liver assist device containing a housing defining a perfusion inlet and a perfusion outlet, a porous membrane structure mounted within said housing to define a perfusion compartment and an adjacent hepatocyte compartment, and porcine hepatocytes isolated from a porcine liver by retrograde perfusion.

Abstract: Disclosed 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 new 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: Methods and compositions for a native-state histoculturing system for skin are disclosed. Skin samples are placed on an extracellular support matrix immersed in a medium whereby the internal surface of the skin is adjacent to the matrix and the external surface is exposed above the surface of the medium and the skin maintained in the medium under skin culturing conditions. An extracellular support matrix comprising a collagen-containing gel and homopolysaccharide sponge or a combination matrix of a top layer of a collagen-containing gel and a bottom layer of a homopolysaccharide sponge is also disclosed. Skin toxicity, hair growth, anti-aging and anti-wrinkling assays utilizing the histoculturing system of the present invention are also disclosed.

Abstract: The present invention relates to a three-dimensional cell and tissue culture system. In particular, it relates to this culture system for the long term culture of liver cells and tissues in vitro in an environment that more closely approximates that found in vivo. The culture system described herein provides for proliferation and appropriate liver cell maturation to form structures analogous to tissue counterparts in vivo. The resulting liver tissues survive for prolonged periods, perform liver-specific functions, and maintain hepatic tissue architecture following in vivo implantation.The liver cultures have a variety of applications ranging from transplantation or implantation in vivo, to screening cytotoxic compounds and pharmaceutical compounds in vitro, to the production of biologically active molecules in "bioreactors" and to the construction of extracorporeal liver assist device.

Abstract: An apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing heart valves is disclosed. Specifically, the present invention relates to an apparatus and method for dynamically seeding and culturing heart valves with human cells, resulting in a heart valve populated with viable human cells. In a preferred embodiment, the apparatus includes a fluid reservoir, a pneumatic pressure chamber, a pressure source for providing a varying pressure to the pressure chamber, and a heart valve holder. In an alternative exemplary embodiment, the apparatus includes a fluid reservoir, a pump, a venturi tube, a heart valve chamber, a heart valve holder, and a valve connected to a timer. In each embodiment, the components are combined in a fluid circuit to create a variable cycling and pulsatile fluid flow for seeding and culturing which can closely resemble the physiological conditions found in the human heart.

Abstract: An apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing vascular grafts is disclosed. Specifically, the present invention relates to an apparatus and method for seeding and culturing vascular grafts with human cells. The apparatus includes a fluid reservoir, a pump, an alternating pressure source, and at least one treatment chamber. By alternating pressure to a support structure within the treatment chamber upon which a vascular graft scaffold is positioned, a varying radial stress is placed on the scaffold. In an alternative embodiment, fluid is pumped directly through the vascular graft subjecting the vascular graft to radial and shear stresses. Applying shear and/or radial stresses to the vascular graft during seeding and culturing simulates physiological conditions.

Abstract: An apparatus and method for the in vitro culturing of animal cells, e.g., mammalian cells, to attain mass growth of the cells per se and/or to recover cell-secreted products of interest therefrom, in which an integral culture and gassing element is formed from spirally-wound or concentricallywrapped lengths of a liquid-impermeable, oxygen-permeable membrane envelope such that narrow cell culture spaces are formed between adjacent windings or wrapping of the envelope. Cells in the culture spaces, which spaces may optionally contain cell substrate material, are cultured in contact with a culture medium introduced into the spaces from an end of the element, and in contact with oxygen-containing gas supplied to the membrane envelope for diffusion across its oxygen-permeable walls into the culture spaces.

Abstract: An apparatus for the large scale culturing and packaging of cell suspensions, three dimensional tissue, and other biological systems is disclosed. The apparatus includes a plurality of flexible or semi-flexible treatment chambers comprising one or more individual culture pockets, a plurality of rigid spacers, an inlet fluid manifold, an outlet fluid manifold, a fluid reservoir, and a means for transporting fluid within the system. During treatment, liquid media is transported from the fluid reservoir to the inlet manifold, which will in turn evenly distribute the media to each of the connected treatment chambers and internal culture pockets. An outlet fluid manifold is also provided to ensure that each treatment chamber is evenly filled and to ensure that any air bubbles formed during treatment are removed from the treatment chambers. The treatment chambers are flexible or semi-flexible so as to provide for easy end-user handling during rinsing and application of the cultured transplants.

Abstract: The present invention provides a medium-penetrating culture carrier comprising a plurality of natural or synthetic threads or the woven body thereof, a method for adhering cells onto this carrier to allow them to be proliferated, and a device which is physically connected to the carrier for feeding a medium using this medium-penetrating culture carrier, which are able to culture three-dimensionally animal cells in order that they can effect self-assembly as they do in the living tissue or organ from which they are derived.

Abstract: Disclosed is a coating compositions for culturing animal adhesive cells comprising a water-insoluble polymer dissolved in a lower alcohol or an aqueous lower alcohol which enable to enhance the adhesive ability and growth of the adhesive cells. Also disclosed is serum-free cell culturing method using culture vessels or carrier coated with the water-insoluble polymer having cell adhesive activity on at least a part of the surface which enable to not only culture but also subculture a variety of adhesive cells including vascular endothelial cell under serum-free condition.

Abstract: The present invention relates to a three-dimensional cell and tissue culture system. In particular, it relates to this culture system for the long term culture of liver cells and tissues in vitro in an environment that more closely approximates that found in vivo. The culture system described herein provides for proliferation and appropriate liver cell maturation to form structures analogous to tissue counterparts in vivo. The resulting liver tissues survivo for prolonged periods, perform liver-specific functions, and maintain hepatic tissue architecture following in vivo implantation. The liver cultures have a variety of applications ranging from transplantation or implantation in vivo, to screening cytotoxic compounds and pharmaceutical compounds in vitro, to the production of biologically active molecules in "bioreactors" and to the construction of extracorporeal liver assist device.

Abstract: An anionic macromolecular containing composition is disclosed that is capable of supporting conservation and differentiation of long-term bone marrow culture initiating cells in cultured mammalian hematopoietic cells. The composition disclosed is a mixture of glycoproteins, more specifically proteoglycans, which are of a molecular weight greater than 200 kD.

Abstract: Method and apparatus for growing cell cultures consisting of a plurality of layers includes a chamber for submerging cells in a medium while supporting and binding them to a semi-permeable membrane. A stirrer is positioned within the chamber and continuously stirs the medium to constantly circulate it past the exposed surface of a semi-permeable membrane on one side of the cell culture and along the exposed surface of the cell culture growing on the membrane and remote from the membrane to transfer nutrients to both surfaces of the cell either directly or through the membrane without disrupting the cell culture on the membrane.