Abstract: The present invention relates to biodetectors for detecting and quantifying molecules in liquid, gas, or matrices. More specifically, the present invention relates to biodetectors comprising a molecular switching mechanism to express a reporter gene upon interaction with target substances. The invention further relates to methods using such biodetectors for detecting and quantifying selected substances with high specificity and high sensitivity.

Abstract: A bio-reactor for enhancing the biomass yield using a closed sterile vessel having a medium, said reactor comprising a central shaft, a support matrix oriented horizontally and dividing the vessel into an upper and a lower chamber, a means for stirring selected from turbine-2,4,6-impeller; marine blade impeller; helical blade impeller; mounted onto the central shaft and located at a predetermined distance from the support matrix so as to generate both radially and axially directed volumetric flow of the media; and at least two semi-circular sparges located both in the upper and lower compartments of the vessel at a predetermined distance from the support matrix.

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 invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties. Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.

Abstract: A process and device are disclosed for simultaneously cultivating different mammal cells, for separately obtaining different mammal cell products and for simulating organic interactions on the humoral plane. Essentially, the invention consists of arranging several culture vessels in a common supply circuit and or cultivating different mammal cells in separate vessels.

Abstract: Improved recombinant retrotransposon vectors for gene transfer are disclosed. The synthetic vectors are truncated so as to reduce or altogether eliminate homologous recombination with retroviral helper sequences found in helper cells used to propagate the vectors, making them safer for use in humans and providing more space for therapeutic genes. The vectors transmit foreign DNA efficiently, are stable, enable abundant RNA expression from the retrotransposon transcriptional promoter, and through their diversity permit many useful applications in therapeutics and transgenics. Methods are described for rescuing tissue-specifics promoters obtaining expression in primary cells, mapping the genome and other techniques of therapeutic and transgenic utility.

Abstract: The present invention provides a module for cell culture comprising a structure comprising hollow fibers and at least two spacers, wherein the spacers have small pores regularly arranged therein and the hollow fibers pass through the small pores and are arranged regularly at a very small distance, a hybrid artificial liver module wherein hepatocytes are immobilized in this module, and a method of cell culture wherein cells are immobilized in the lumen or extra-fiber space of the module of this module by the use of a centrifugal force and wherein the immobilized cells are cultured. The present invention also provides a method for cell culture wherein hepatocytes are cultured in a culture medium containing DMEM as a basal medium.

Abstract: Membrane module and process for its manufacture, wherein the membrane module contains at least two groups of hollow-fiber membranes, the groups arranged in layers and capable of being fed independently by fluids. The membrane module has a housing shell comprising a channel-shaped housing middle section, open at the top, and adjoining end pieces, also open at the top and with a number of arms corresponding to the number of groups. The groups of hollow-fiber membranes are inserted as layers over one another such that they are arranged in the direction of the longitudinal extent of the housing middle section and substantially parallel to each other. The ends of hollow-fiber membranes of different groups are arranged in respectively different arms of the end pieces and embedded with a sealing compound such that at least one end of the hollow-fiber membranes is open.

Abstract: The invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties. Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.

Abstract: A bioreactor (10) has arranged within its internal chamber (14) a bundle of liquid-impermeable hollow tubes (16), which are used to freeze or vitrify a biologically active material seeded within the internal chamber. When the bioreactor is ready for use, the biologically active material may be thawed by perfusing the liquid-impermeable hollow tubes with a heated solution or heated vapor.

Abstract: HCMV glycoproteins B and H have been identified. The gB protein is encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary. The gH protein is encoded by DNA in the HindIII L fragment lying between 228 and 2456 bases from the L/D boundary.The genes have been incorporated in recombinant vaccinia vectors and expressed in host animals to raise HCMV-neutralising antibody, thereby indicating vaccine potential. The glycoproteins can also be used in a variety of different ways, as vaccines or in the production, purification or detection of HCMV antibody.

Abstract: A novel cell seeded hollow fiber bioreactor is described as a potential bioartificial kidney. Endothelial cells along with pericyte, vascular smooth muscle, and/or mesangial cells or any mesenchymally derived support cells are seeded along a hollow fiber in a perfused bioreactor to reproduce the ultrafiltration function and transport function of the kidney. Maintenance of tissue specific function and ultrastructure suggest that this bioreactor provides an economical device for treating renal failure.

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: The invention relates to a method of removing an agent from a suspension of cells using a semi-permeable membrane. In one aspect of the invention, the cells are used to bioprocess a biological fluid after removal of the agent.

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: The invention features an empty device for receiving a bioactive agent. The device includes a biocompatible and semi-permeable membrane that defines an enclosed space; the membrane also has at least one end that defines an opening for introducing the bioactive agent into the enclosed space. The device is configured to be placed in an animal.In one embodiment of the invention, the membrane has an inner surface and an outer surface, where the inner surface defines the inner surface, and includes a biocompatible adhesive in the general region of the opening to allow sealing of the opening after the introduction of the bioactive agent into the enclosed space.Another embodiment of the invention includes a biocompatible frame mounted in supporting relationship to the membrane and defining an opening for introducing the bioactive agent into the enclosed space. The frame has greater porosity than the membrane.

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: A "micro" hollow fiber bioreactor and method of use are provided for use in screening different cell lines and process conditions. The bioreactor includes the use of an oxygen permeable (e.g., silicone rubber) tube sealably containing a hollow fiber bundle, in order to create an extracapillary space to provide a medium reservoir and an intracapillary space for the growth of cells. The bioreactor avoids the need for oxygen or medium pumps or supply systems, and permits multiple cell lines, and/or multiple conditions to be evaluated simultaneously. Preferably, the tube has an oxygen permeability of between about 100.times.10.sup.-10 to about 10,000.times.10.sup.-10 (cc-mm/sec-cm.sup.2 -cm Hg), the extracapillary space provides a medium reservoir of about 1 ml to about 100 ml, the intracapillary space provides a cell culture volume of about 0.1 ml to about 1 ml, the hollow fibers have a molecular weight cut off from about 1 kD to about 1,000 kD and a pore size of from about 0.

Abstract: Improved hollow fiber bioreactor systems and cell culture methods are described. The improvement includes means which may be a novel basket bioreactor for extending the extracapillary space of a conventional hollow fiber bioreactor.

Abstract: Methods and apparatus for the extraction of DNA from a suspension of cells are described. The methods utilise a hollow membrane filter to separate DNA from cellular debri after lysis of cells. The suspension of cells can be a suspension of cultured cells or cells contained in a body fluid such as blood. An ion-exchange step can be included in methods so that purified DNA is provided. The apparatus (1) comprises a vessl (2) in which cells can be cultured. The vessel has a hollow membrane filter (5) associated with an end thereof. The described methods can be used for extracting genomic DNA from cells but are particularly suitable for extracting plasmid DNA from microorganisms.

Abstract: Methods of maintaining animal cells for product production, for supporting hepatocyte function and viability to treat a patient suffering from hepatic failure and for preserving tissue-specific function of mammalian cells are carried out with a bioreactor containing a feed and waste chamber and a cell chamber separated by a selectively permeable membrane. Within the cell chamber, a biocompatible contracted three-dimensional gel matrix entraps animal cells or genetic modifications thereof, and a liquid phase contains a concentrated solution of the cell product. The bioreactor uses only two chambers to achieve three distinct zones within the bioreactor. The bioreactor can be of either hollow fiber or flat-bed configuration. In the configuration using hollow fibers, the two fluid paths correspond to the cavity surrounding the hollow fibers (the extracapillary space), and to the lumens of the hollow fibers themselves. Both fluid paths have inlet and outlet ports.

Abstract: A macroencapsulation device for somatic cells using ultrapurified Na alginate and polysulfone hollow fibers of 30 kDa molecular weight cutoff. Ultrapurified Na alginate material is used which has a high `G` content, low endotoxin content, low divalent metal toxins and low protein impurities. Islet cells prior to being encapsulated, are irrigated with Hank's modified solution (without Ca and Mg) containing gentamycin, vancomycin and amphotericin B and then passed through a leukoabsorb filter to reduce the donor antigen load of passenger leukocytes and to reduce the bioburden of microorganisms including viruses. Encapsulation is done in RPMI 1640 tissue culture fluid containing necessary nutritional supplements and ATP source of energy. The open ends of the fiber are covered with a porous membrane. To further improve biocompatibility, the outer wall of the polysulfone is lightly gelled with alginate gel.

Abstract: A bioreactor containing living animal cells at a density approaching that of normal animal tissue is described. High cell loading is achieved by providing a flow restrictor which controls fluid flow through the bioreactor during cell loading. Methods for making and using the bioreactor are also described.

Abstract: Cells such as hepatocytes are applied to hollow fibers by forming a first gelled matrix layer on the outside surface of the fibers, adhering cells to the matrix layer and forming a second gelled matrix layer covering the adhered cells. To form the matrix layers, a liquid matrix-forming material such as containing collagen is applied and then gelled. An acidic liquid matrix-forming material may be used to prepare the second matrix layer. The hollow fibers may be cooled to less than 15.degree. C. before forming the first gelled matrix layer. Three-dimensional co-cultures may be formed by applying non-parenchyma cells to the inside of the walls of the hollow fibers.

Abstract: This invention relates to methods and compositions of controlling cell distribution within a bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the BAO with ECM molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination.

Abstract: The invention relates to an implant obtained by assembling in vitro various elements in order to form a neo-organ which is introduced preferably in the peritoneal cavity of the recipient. The implant comprises a biocompatible support intended to the biological anchoring of cells; cells having the capacity of expressing and secreting naturally or after recombination a predetermined compound, for example a compound having a therapeutical interest; and a constituent capable of inducing and/or promoting the gelling of said cells. The invention also relates to a kit for the preparation of the implant as well as to a new recombinant retroviral vector comprising a provirus DNA sequence modified in that the genes gag, pol and env have been deleted at least partially so as to obtain a proviral DNA capable of replication. The invention also relates to recombinant cells comprising the new retroviral vector.

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: The invention is directed to methods of culturing rickettsiae in Ixodes scapularis cell lines. The methods of the invention provide for culture of microorganisms such as Anaplasma marginale, Ehrlichia canis, and Rickettsia rickettsii. A method of the invention involves incubating a rickettsia with an Ixodes scapularis tick cell culture in a culture medium under reduced oxygen and increased CO.sub.2 at a sufficient temperature until growth of the rickettsia is detected. The culture medium comprises a medium suitable for the growth of invertebrate cells supplemented with an organic buffer. The cell culture method can be used in large scale production of rickettsia containing products useful in diagnostic assays and vaccine preparations.

Abstract: Continuous cultures of pluripotent parenchymal hepatocytes were derived from the epiblasts of pig blastocysts. The cultures are feeder-dependent and grow slowly with doubling times of 3 to 4 days. They differentiate into large secretory duct-like structures or form small canaliculi. In combination with feeder cells and, optionally, adult pig hepatocytes and macrophages, the cells are useful in an artificial liver device which may be utilized as temporary liver support for the mitigation of the pathological effects of liver failure.

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: This invention relates to methods and compositions of controlling cell distribution within a bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the BAO with ECM molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination.

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: A cell growing device for in vitro cell population growth includes at least one hollow fiber cartridge having a plurality of capillaries at least one of which is selectively permeable. The flow of media out of a lumen of the cartridge is substantially blocked off thereby forcing media flowing into the lumen via an inflow opening to permeate across the capillaries of the cartridge and into the extracapillary space thereof.

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 unique hollow fiber multiple bioreactor system and method for the propagation of cells and the production of various cell propagation products is described. Simultaneous fluid flow through lumens of hollow fibers in bioreactors is maintained constant across multiple bioreactors so production is maintained essentially constant. The system and method include use of a flow block holder which allows a dissolved oxygen probe to be calibrated at the same pressure as the media in the bioreactor system.

Abstract: An immunotherapy protocol in which leukocytes are cultured in the presence of interleukin-2 in a hollow fiber cartridge perfusion system for at least four days to achieve a harvest yield of at least 100% of leukocytes with respect to the number of cells initially present in the hollow cartridge, and where the leukocytes have a lytic activity at least equal to that of cells grown in a static culturing system and where the lytic activity of the cells following the culture period in the hollow fiber is at least equal to cells grown in a static culturing system.

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: A method for maintaining hepatocytes in culture includes providing the hepatocytes with a support, the support including extracellular matrix, the support having a configuration that permits each of at least a portion of the hepatocytes to form at least one apical surface and at least two discrete basal surfaces.