Abstract: A method of inducing the proliferation and/or differentiation of human adult pancreatic cells entails contacting primary cultures of such cells with Hepatocyte Growth Factor/Scatter Factor (HGF/SF), thereby inducing a proliferation of .beta.-epithelial cells, an increase in the number of .beta.-epithelial cells which form islet-like cell clusters, and an increase in insulin production per cell. The method is improved by culturing the cells on an extracellular matrix such as 804G in the presence of HGF/SF, and is further improved by reaggregating thus-treated cells and contacting said cells with an insulin gene upregulating agent such as a poly(ADP-ribose) synthetase inhibitor such as a nicotinamide or benzamide. The method provides increased numbers of functional islet-like cell clusters for transplantation.

Abstract: The present invention describes a multi-well bone cell culture device for use in the quantitative and/or qualitative assessment of bone cell activity, the device comprising a flat base having a sintered film of calcium phosphate entities on one side and an open-ended multi chamber unit positioned on top of the film which by a sealing means is sealed to the film coated base forming individual containment wells. The device is useful in the analysis of bone cell function, for the screening and determination of bone cell disease, and the development of drugs to alter bone cell activity.

Abstract: Methods and artificial matrices for the growth and implantation of urological structures and surfaces are disclosed in which urothelial cells are grown in culture on biodegradable, biocompatible, fibrous matrices formed of polymers, such as polyglycolic acid, polylactic acid, or other polymers which degrade over time. The cells can be cultured in vitro until an adequate cell volume and density has developed for the cells to survive and proliferate in vivo. Alternatively, when adequate cell numbers for implantation are available, the cells can be attached to the matrix and implanted directly, without proliferation in vitro. The implants approximate the desired urological structure to be replaced or repaired, such as the kidney, urether, bladder, urethra, and the like. Implantation is followed by remodeling through cell growth and proliferation in vivo.

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: A device having a surface coated with a firmly adherent cell monolayer is produced by culturing adherent cells in the presence of the surface in vitro under conditions of continuous shear stress of from 0.4 dyne/cm.sup.2 to 33 dyne/cm.sup.2 produced by the force of circulating fluid medium in contact with the cells. The surface may be contained by an implantable device, or a culture or fermentation vessel. Preferably, an endothelial cell monolayer is produced on a surface of a prosthetic vascular device made of polypropylene. In a hollow fiber cartridge device, endothelial cells are grown under shear stress on the inner surface of the lumen of a hollow fiber and perivascular cells are grown on the outer surface of the fiber. Growing cells under continual stress more closely approximates the in vivo environment where blood passes over the endothelium in a blood vessel, and produces a cell monolayer closely resembling naturally occurring firmly adherent cell layers found in vivo in the lining of blood vessels.

Abstract: Bone and cartilage are regenerated in a patient by a process of removing fatty tissue such as omentum tissue from a patient, comminuting the tissue to form small tissue particles, suspending the particles in a liquid to form a suspension, depositing the suspension on a solid carrier to prepare a solid implanting material, implanting the implanting material in a patient in an environment favoring bone or cartilage formation, and regenerating bone or cartilage in the patient. The carrier can be demineralized bone, collagen, mineral material or synthetic polymer material in pulverulent, textile, porous particle or monolith form. A cell adhesion agent may be applied to the carrier or added to the suspension, and a growth factor may be deposited on the carrier. Comminuting is performed by digesting with an enzyme and/by mechanically comminuting. Liquid used to form the suspension may contain a gel precursor which is gelled after the suspension is deposited to the carrier.

Abstract: A bioartificial three-dimensional hydrogel extracellular matrix derivatized with a cell adhesive peptide fragment is provided for use in tissue regeneration or replacement. The choice of adhesive peptide fragment depends on the desired target cell type. Cartilage or tendon can be regenerated by implanting a matrix containing adhesive peptide fragments that favor chondrocyte invasion. The matrix can be pre-seeded with cells, and tissue can be reconstituted in vitro and then implanted. A cell-seeded matrix can be encapsulated in a semi-permeable membrane to form a bioartificial organ. An agarose hydrogel matrix having an agarose concentration of 0.5-1.25% (w/v) and an average gel pore radius between 120 nm and 290 nm is preferred. The peptide fragment preferably contains the sequence, ArgGlyAsp or TyrIleGlySerArg or IleLysValAlaVal, and is covalently immobilized to the matrix.

Abstract: A human pre-oligodendroglial stem cell line, HOP-1, is provided herein. The cell line is useful for methods of treatment of central nervous system disorders, as well as for neuropharmaceutical drug discovery.

Abstract: The invention includes methods for the isolation and clonal propagation of mammalian neural stem cells. The methods employ a novel separation and culturing regimen and bioassays for establishing the generation of neural stem cell derivatives. These methods result in the production of non-transformed neural stem cells and their progeny. The invention demonstrates, at the clonal level, the self regeneration and asymmetrical division of mammalian neural stem cells for the first time in feeder cell-independent cultures. Lineage restriction is demonstrated within a developing clone and is shown to be sensitive to the local environment. Multipotent neural stem cells cultured on a mixed substrate of poly-D-lysine and fibronectin generate PNS neurons and glia, but on fibronectin alone the stem cells generate PNS glia but not neurons. The neurogenic potential of the stem cells, while not expressed, is maintained over time on fibronectin.

Abstract: A method is disclosed for the efficient production of a transfected cell which comprises a step of culturing transfected cells in the presence of a cell-adhesive substance, after injection of a foreign gene into target cells, upon production of the transfected cells by transfer of a foreign gene into the target cells through cell perforation. Also provided are transfected cells produced by the method, and a kit for the production of transfected cells that includes a cell-adhesive substance as an essential component, which kit is suitable for use with the method for the efficient production of transfected cells by cell perforation.

Abstract: Methods for producing thin colloidal silica films on substrates, such as corona treated polystyrene, are provided. The dried films are characterized as 50 nm thick, high silanol, homogenous, high surface area, porous, uncracked, adherent, wetting, negatively charged, and gamma radiation stable. Several differential advantages of the colloidal silica films were demonstrated in epithelial cell culture, especially regarding primary cultures in serum free media. Cell responses to the films were increased explant adhesion, increased cell growth rate, and increased expression of differentiated function before and after subculture as compared to tissue culture polystyrene.

Abstract: In vitro production of clinically useful quantities of single species of mature, differentiated human blood cells is carried out by a method in which human pluripotent hematopoietic stem cells, preferably from a universal donor, are incubated in a bioreactor in a growth medium also containing specific combinations of recombinant human growth and maturation promoting polypeptide factors that expand stem cell cultures and promote the maturation and differentiation of stem cells into single species of erythroid, thrombocytic or granulocytic human blood cells, and harvesting the mature cells. The growth and maturation promoting polypeptides employed include SCGF, Interleukins 1,3,4,5,6, and 11, GM-CSF, M-CSF, G-CSF and EPO. Stem cells may be preliminarily genetically modified so as to remove histocompatibility or blood group antigens with which a recipient may be incompatible, or the stem cells may be genetically altered by transfection with appropriate DNA-containing vectors, prior to addition to the bioreactor.

Abstract: Novel non-crystalline, porous bioactive glass and ceramic materials that permit the in vitro formation of bone tissue when exposed to a tissue culture medium and inoculated with cells are disclosed. The present invention also discloses methods of treating bioactive glass materials to control pH so that when the glass is exposed to a tissue culture medium and then inoculated with cells, bone tissue growth occurs in vitro. The glass material disclosed is preferably formed from SiO.sub.2, CaO, Na.sub.2 O and P.sub.2 O.sub.5 and the porous, non-crystalline structure is most preferably created by melting the constituents, cooling and pulverizing the resulting glass, and then forming and hot pressing the powder. The glass of the present invention may be formed to produce templates that are useful for various indications, as well as granules that may be formed into a paste.

Abstract: A matrix structure containing attached cells such as endocrine cells, fibroblasts, endothelial cells or genitourinary cells is implanted in a patient adjacent tissue having a high surface area and vasculature such as mesentery, omentum or peritoneum tissue. Large volumes of cells can be attached to the matrix and the matrix implanted with minimum trauma and blood loss into a patient to produce a functional organ equivalent. Multiple matrix structures containing cells can be implanted to functionally resemble naturally occurring organs. Implanting multiple matrices between folds of the mesentery is particularly well suited for growth of endocrine structures, including liver, pancreas, and adrenal gland. The matrix structure is preferably formed from a biodegradable artificial polymer. Collagen and non-biodegradable materials can also be used, and the matrix structure can be overlaid with a material that enhances cell attachment.

Abstract: In an apparatus for culturing cells which comprises a plate-like body with a lattice structure having openings separated from one another by sidewalls and a bottom disposed on one side of the lattice structure and having passages permeable for liquids but not for cells so as to form, with the lattice structure, cavities for receiving cells to be grown therein, the cavities have a clear width of between 50 .mu.m and 1000 .mu.m and the bottom consists of a material to which cells placed into the cavities will not easily attach so that the cells grow from the side walls of the cavities causing them to form a three-dimensional cell structure.

Abstract: A biocompatible capsule for containing cells for implantation is prepared containing an inner support that provides tensile strength to the capsule. The capsule may be a tubular semipermeable membrane such as a hollow fiber membrane having both ends sealed. A rod shaped inner support extends through the lumen and ends of the rod are attached to sealed ends of the fiber. Prior to sealing one fiber end, cells are introduced into the lumen. Cells within the capsule may be suspended in a liquid medium or immobilized in a hydrogel or extracellular matrix material, and biologically active molecules can be delivered from the capsule to surroundings or from the surroundings into the capsule. The inner support may have external features such as flutes or a roughened or irregularly-shaped surface, and may be coated with cell-adhesive substance or a cell-viability-enhancing substance.

Abstract: Provided are methods and compositions for the repair of articular cartilage defects in a mammal. Denuded chondrogenic cells are proliferated ex vivo as monolayer cultures in order to expand the pool of available chondrogenic cells. During proliferation the chondrogenic cells stop secreting the extracellular matrix components, type II collagen and sulfated proteoglycans. The proliferated cells then are seeded into a pre-shaped well having a cell contacting, cell abhesive surface. The cells cultured in the well redifferentiate and begin to secrete cartilage-specific extracellular matrix again. Accordingly, essentially unlimited amounts of synthetic cartilage may be prepared from small samples of biopsy tissue. Also provided are methods for surgically repairing articular cartilage defects in mammals using the synthetic cartilage prepared in accordance with the invention.

Abstract: A cell-scaffold composition is prepared in vitro for implanting to produce functional organ tissue in vivo. The scaffold is three-dimensional and is composed of hollow or solid fibers of a biocompatible, synthetic polymer which is biodegradable or non-biodegradable. The fibers of the scaffold may have a branched configuration extending outwardly from a central stem. Fibers of the scaffold are spaced apart such that the maximum distance over which diffusion of nutrients and gases must occur through a mass of cells attached to the fibers is between 200 and 300 microns. The diffusion provides free exchange of nutrients, gases and waste to and from cells proliferating throughout the scaffold in an amount effective to maintain cell viability throughout the scaffold in the absence of vascularization.

Abstract: The isolated proteins kalinin and k-laminin are disclosed to provide adhesion between epidermal keratinocytes and the underlying dermis. Purified kalinin localizes to the anchoring filaments of basement membranes of human subepithelial skin, trachea, esophagus, cornea and amnion when such areas are probed with BM165 monoclonal antibody after localization. The protein has a molecular weight of approximately 410-495 kDa and exists in a cell-associated form (about 495 kDa) and two medium-associated forms (about 460 and 410 kDa, respectively). The BM165 epitope is located on the 165-kDa and 200 kDa subunits. Kalinin has a rotary-shadow image revealing an asymmetric rod 107-nm long having two globules at a first end and a single globule at an opposing end. The k-laminin adhesion molecule is an isolated heterotrimeric laminin variant that has a molecular weight of about 650 kDa and separates on western blots into first and second fragments that are similar to the B1 and B2 fragments of laminin.

Abstract: Fibers of a biocompatible, biodegradable or non-biodegradable, synthetic polymer are provided, and are formed into a three-dimensional scaffold. The fibers of the scaffold may have a branched configuration extending outwardly from a central stem. The fibers provide sufficient surface area to permit attachment to the scaffold in vitro of an amount of cells effective to produce functional vascularized organ tissue in vivo. Fibers of the scaffold are spaced apart such that the maximum distance over which diffusion of nutrients and gases must occur through a mass of cells attached to the fibers is between 200 and 300 microns. The diffusion provides free exchange of nutrients, gases and waste to and from cells proliferating throughout the scaffold in an amount effective to maintain cell viability throughout the scaffold in the absence of vascularization.

Abstract: A composition and method are provided for inhibition of vascular smooth muscle cell proliferation following injury to the endothelial cell lining of a blood vessel such as resulting from angioplasty, vascular bypass surgery or organ transplantation. The composition is a matrix such as a biodegradable hydrogel made of a synthetic polymer, protein or polysaccharide seeded with vascular endothelial cells which can be xenografts, allografts or autografts, or genetically engineered cells. Attachment of cells to the matrix can be enhanced by coating with collagen, laminin, fibronectin, fibrin, basement membrane components or attachment peptides. Biologically active compounds such as anti-inflammatory agents may also be contained in the matrix. In the method, the matrix containing endothelial cells is implanted in a patient at a site adjacent the injury such as by wrapping the matrix around the blood vessel.

Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.

Abstract: A method for producing a neuroblast and a cellular composition comprising an enriched population of neuroblast cells is provided. Also disclosed are methods for identifying compositions which affect neuroblasts and for treating a subject with a neuronal disorder, and a culture system for the production and maintenance of neuroblasts.

Abstract: Methods, compositions and devices are provided for the growth of human stem and/or hematopoietic cells in culture. Bioreactors are provided in which diverse cell types are simultaneously-cultured in the presence of appropriate levels of nutrients and growth factors substantially continuously maintained in the bioreactor while removing undesirable metabolic products. This simultaneous culture of multiple cell types successfully reconstructs hematopoietic tissue ex vivo. Optionally, at least one growth factor is provided through excretion by transfected stromal cells, particularly heterologous cells. The stromal cells and hematopoietic cells may be maintained separately, and both the adherent and non-adherent cells harvested.

Abstract: A cell-scaffold composition is prepared in vitro for implanting to produce functional organ tissue in vivo. The scaffold is three-dimensional and is composed of fibers of a biocompatible, biodegradable, synthetic polymer. Cells derived from vascularized organ tissue are attached in vitro to the surface of the fibers uniformly throughout the scaffold in an amount effective to produce functional vascularized organ tissue in vivo. Fibers of the scaffold are spaced apart such that the maximum distance over which diffusion of nutrients and gases must occur through a mass of cells attached to the fibers is between 100 and 300 microns. The diffusion provides free exchange of nutrients, gases and waste to and from cells proliferating throughout the scaffold in an amount effective to maintain cell viability throughout the scaffold in the absence of vascularization.

Abstract: A tissue graft construct comprising submucosal tissue of a warm-blooded vertebrate and a proliferating population of cells is described. The submucosal graft constructs used in accordance with the present invention induce host tissue proliferation, remodeling and regeneration of appropriate tissue structures upon implantation in a host.

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: Methods and artificial matrices for the growth and implantation of cartilaginous structures and surfaces and bone are disclosed. In the preferred embodiments, chondrocytes are grown on biodegradable, biocompatible fibrous polymeric matrices. Optionally, the cells are proliferated in vitro until an adequate cell volume and density has developed for the cells to survive and proliferate in vivo. One advantage of the matrices is that they can be cast or molded into a desired shape, on an individual basis, so that the final product closely resembles a patient's own ear or nose. Alternatively, flexible matrices can be used which can be manipulated at the time of implantation, as in a joint, followed by remodeling through cell growth and proliferation in vivo. The cultured cells can also be maintained on the matrix in a nutrient media for production of bioactive molecules such as angiogenesis inhibiting factor.

Abstract: Anchorage-dependent cells are grown on a novel substratum which is believed to increase the oxygenation of the cells and reduce the formation of free-radicals. The substratum consists of a perfluorocarbon reservoir which is coated with a layer of protein (e.g.,collagen or gelatin) which has been perfluoroalkylated. The perfluoroalkylated protein forms a very stable surface to which anchorage-dependent cells can attach and grow. Using this system, mammalian cell cultures have been grown to densities exceeding 10.sup.7 cells/cm.sup.2. The increased density is attributed to the formation of multiple cell layers which resemble tissue-like structures.

Abstract: The peptide X-Arg-Gly-Asp-R-Y wherein X is H or at least one amino acid and Y is OH or at least one amino acid, and R is an amino acid selected from Thr or Cys, or other amino acid, having the same cell-attachment activity as fibronectin and the peptide X-Arg-Gly-Asp-Ser-Y, wherein X and Y, having said activity are disclosed.

Abstract: The present invention provides an artificial organ system comprising an endothelial cell layer, an epithelial cell layer and an artificial microporous membrane disposed between and indirect contact with the endothelial cell layer and epithelial cell layer such that the membrane has an endothelial side and an epithelial side.

Abstract: Animal adhesive cells, particularly human vascular endothelial cells, are cultured in serum-free condition by coating at least one polymer having cell adhesive activity on an inner surface of a culture vessel or surface of a carrier for cell culture, and culturing the cells in the coated vessel or with the coated carrier using a serum-free medium for animal cell culture containing isolated serum albumin, and preferably also transferrin. The polymer is a synthetic polymer modified with a peptide having cell adhesive activity or a natural polymer having cell adhesive activity or a combination thereof. Preferably, the peptide is RGDV, RGDS, RGDN, DGEA or YIGSR and the natural polymer is collagen, gelatin, keratin, fibronectin, vitronectin or laminin. A preferred medium for culturing human vascular endothelial cells is basal medium MCDB 131 or MCDB 107 containing isolated serum albumin, transferrin, hydrocortisone and epithelial growth factor.

Abstract: A method of growing complete vertebrate skin in vitro, which comprises obtaining a segment of vertebrate skin, positioning the skin segment in an artificial cell-growth medium containing sufficient nutrients to maintain growth of cells of the skin, and subjecting the skin segment to stretching forces while the skin segment is in the medium. Skin produced by the method and an apparatus for carrying out the method are also part of the present invention.

Abstract: A method of increasing the number of adult pancreatic islet cells available for transplantation by contacting the cells with laminin 5 extracellular matrix. When contacted with the deposited matrix produced by 804G rat bladder carcinoma cells, a 1,500 fold increase in cell number is observed after three passages in culture. Islet cell expansion also occurs when cells are contacted with 804G soluble matrix. The expanded islet cells contain insulin and respond to glucose challenge.

Abstract: A method of expanding the number of pancreatic islet cells for transplantation. Fetal islet cells are cultured in the presence of laminin 5 extracellular matrix, resulting in a significant increase in cell number after passaging in culture. The expanded islet cells contain insulin and respond to glucose challenge.

Abstract: The present invention features a device and method for inducing cell differentiation. One embodiment of the present invention features a device comprising a wall having an interior surface and an exterior surface. The interior surface defines a chamber. The exterior surface is capable of being placed in contact with biological tissues. The wall has an opening in communication with the chamber for receiving an induction material and for closing the chamber to contain the induction material. The wall comprises a biologically compatible, permeable material capable of receiving cells capable of differentiation. The device is used to contain an induction material in a biological environment and receiving cells in the chamber. The cells are allowed to differentiate in the presence of the induction material.

Abstract: The present invention provides a method of growing smooth muscle cells in a host with the purpose of maintaining natural cellular function by the function of smooth muscle tissue. In this method, smooth muscle cells are freed from isolated smooth muscle tissue then cultured and injected into the host in combination with an extracellular matrix.

Abstract: Novel non-crystalline, porous bioactive glass and ceramic materials that permit the in vitro formation of bone tissue when exposed to a tissue culture medium and inoculated with cells are disclosed. The present invention also discloses methods of treating bioactive glass materials to control pH so that when the glass is exposed to a tissue culture medium and then inoculated with cells, bone tissue growth occurs in vitro. The glass material disclosed is preferably formed from SiO.sub.2, CaO, Na.sub.2 O and P.sub.2 O.sub.5 and the porous, non-crystalline structure is most preferably created by melting the constituents, cooling and pulverizing the resulting glass, and then forming and hot pressing the powder. The glass of the present invention may be formed to produce templates that are useful for various indications, as well as granules that may be formed into a paste.

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: A cell culture apparatus includes a substrate, and an inorganic film on the substrate and having a relatively smooth surface for cell growth thereon. Moreover, the relatively smooth surface has a relatively low predetermined surface roughness so that cells grown thereon adhere by chemical adhesion rather than by mechanical interlocking. In other words, the predetermined surface roughness defines surface indentations smaller than a size of corresponding cells to be grown thereon. Accordingly, the interaction between various inorganic materials and cells may be studied based upon chemical adhesion rather than mechanical interlocking. The predetermined surface roughness is preferably less about 10 .mu.m root-mean-square, and more preferably less than about 1 .mu.m root-mean-square. The relatively smooth growth surface is preferably obtained by thermal vacuum evaporation and deposition, followed by annealing of the as-deposited film.

Abstract: This disclosure includes a method for generating a functional hybrid bioprosthesis. Tissue formed naturally of interstitial collagens is treated to kill native cells and remove potentially immunologically active soluble molecules. Then it may be treated sequentially with extracellular matrix adhesion factor, extracellular matrix glycosaminoglycan, and growth factor appropriate to the cell type required to function within the matrix, and incubating the transplant tissue matrix with cells that are either allogeneic or autologous for the recipient thereby imparting to the matrix the characteristics of the cell type and tissue selected. Tissues with a variety of functional bioactivities can thus be formed in vitro prior to graft transplantation or implantation which will exhibit reduced or no stimulation of an immunological response in the recipient.

Abstract: The present invention provides fibronectin self-assembly sites. The invention provides a set of polypeptides derived from the first type III repeat of fibronectin which contain a fibronectin-fibronectin binding site. These polypeptides have been used to obtain a second set of polypeptides derived from the C-terminal type I repeats which contain a second fibronectin-fibronectin binding site which interacts with the first type III repeat of fibronectin. These polypeptides are capable of inhibiting fibronectin matrix assembly by interfering with fibronectin-fibronectin binding. These polypeptides are also capable of enhancing fibronectin matrix assembly and inducing disulfide cross-linking of fibronectin molecules in vitro. In addition, these polypeptides are capable of inhibiting migration of tumor cells. The polypeptides of the present invention have a number of related uses as well.

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: Hepatocytes spheroids can be formed by culturing hepatocytes in a culture vessel using a lipid-bound glycosaminoglycan as a culture substrate. Floating spheroids of hepatocytes can be obtained efficiently, which are capable of maintaining liver-specific functions and of keeping the spheroid form stably for a prolonged period of time.

Abstract: A high performance hollow fiber bioreactor having concentric hollow fiber bundles: a central hollow fiber bundle supplies media, and an outer array supplies oxygen needed for cell culture. Useful to expand therapeutic cells such as stem cells ex vivo, and as an extracorporeal device such as an artificial liver.

Abstract: This disclosure includes a method for generating a functional hybrid bioprosthesis. Tissue formed naturally of interstitial collagens is treated to kill native cells and remove potentially immunologically active soluble molecules. Then it may be treated sequentially with extracellular matrix adhesion factor such as fibronectin, extracellular matrix glycosaminoglycan such as heparin, and growth factor appropriate to the cell type required to function within the matrix, and incubating the transplant tissue matrix with cells that are either allogeneic or autologous for the recipient thereby imparting to the matrix the characteristics of the cell type and tissue selected. Tissues with a variety of functional bioactivities can thus be formed in vitro prior to graft transplantation or implantation which will exhibit reduced or no stimulation of an immunological response in the recipient.

Abstract: The substrate for cell culture to be used for cell arrangements is formed by applying a photoresist on a surface of a substrate, removing selective parts of the photoresist on the surface of the substrate by optical exposure and development, and forming an immobilized enzyme membrane on the surface of the substrate after removing the photoresist, and removing the photoresist after forming the immobilized enzyme membrane. An enzyme substrate of enzyme contained in the immobilized enzyme membrane is a material that is necessary for growth of cells for forming cell arrangements or is a material that inhibits growth of such cells, and a reaction product of oxygen contained in the immobilized enzyme membrane is a material that is necessary for growth of cells for forming cell arrangements or a material that inhibits growth of such cells. It is possible to control the cell adhesion on the surface of the substrate.