Abstract: Aggregates and their method of preparation suitable for implantation into a recipient in order to produce insulin in vivo. The methods involve culturing islet cells isolated from the pancreas of donor piglets with isolated Sertoli cells from the testes of donor piglets. A preferred period of culturing is 5 days and may be followed by a purification procedure.

Abstract: The present invention relates to modified cells carrying a heterologous gene sequence encoding a protein, such as an Inhibitor of differentiation (Id) gene sequence that binds a basic helix-loop-helix (bHLH) protein to inhibit cell growth, differentiation and/or tumorigenesis of the modified cells. The modified cells are differentiated, proliferate and do not become tumorigenic when grafted into a recipient subject. Additionally, the modified cells produce a factor or factors that enhance the viability of co-grafted organs, tissues or cells. Thus, the modified cells are useful for testing agents for effects on the cells, for co-grafting with transplant organs, tissues or cells. The modified cells are also useful for enhancing the viability of thawing cells that have been cryo-preserved. In one embodiment, the modified cells are modified Sertoli cells.

Abstract: The invention provides cell culture devices comprising a channel, the channel comprising one or more inlets and one or more outlets, and a cell retention chamber defined by an internal surface of the channel and a plurality of projections extending therefrom. The invention further provides methods of use relating to such cell culture devices.

Abstract: The present disclosure covers compositions and method for the preparation and use of mixtures of adult stem/progenitor cell populations recovered and enriched from specific tissues with very limited attempts for their purification. Such mixtures of cell populations have improved therapeutic effectiveness in the treatment of certain diseases and tissue regeneration treatments over their more purified counterpart cell populations. Such mixtures of cell populations can be cryopreserved for future clinical use.

Abstract: A method for promoting the in vitro multiplication of human melanocytes and/or the freezing thereof, notably human melanocytes obtained from individuals of phototype IV, V or VI or from hyperpigmentary lesions entails introducing into a culture of same, a thus effective amount of at least one 1-phenyl-3-(2-thiazolyl)-2-thiourea (PTU) compound or PTU analogue or PTU mimetic selected from the group consisting of vitamin C, arbutin, hydroquinone, kojic acid or acid or ester derivative thereof, ellagic acid, aminophenol derivative, procysteine or derivative thereof, niacinamide, isothiocyanate, thiocyanate, lucinol and mixtures thereof; also provided thereby can be melanocyte libraries, co-cultures and reconstructed epidermides and/or reconstructed skin that are highly pigmented which are useful for the study of pigmentation disorders, for the screening of cosmetic or dermatological active agents, or for the treatment of skin lesions, in particular in individuals with a high phototype.

Abstract: A cell culture system related to extended in vitro culture of mature neuronal cells and methods for preparing the cell culture system are provided. In a preferred embodiment the invention provides a cell culture system comprising a mixture of mature neuronal retinal cells and cells isolated from a ciliary body. Methods for identifying bioactive agents that alter neurodegeneration of neuronal retinal cells are also provided.

Abstract: The subject invention is directed to a mixed cell composition to generate a therapeutic protein at a target site by providing a first population of mammalian cells transfected or transduced with a gene that is sought to be expressed, and a second population of mammalian cells that have not been transfected or transduced with the gene, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.

Abstract: A process for identifying compounds that can modulate the release of neuromediators is described in which, for example, at least one compound that is to be tested is brought into contact with a nerve tissue preparation and the possible modulating effect of the compound on release of neuromediator by the nerve tissue preparation is determined. Methods of preparing calibrated pieces of mammalian cerebral material and kits for the implementation of the process are also described.

Abstract: Tissue engineered constructs including a matrix and cells transfected with a gene for a growth factor. The constructs may be implanted into a tissue site, where the growth factor gene enhances a metabolic function furthering integration of the construct in the tissue site. If the matrix is biodegradable, the metabolic result may include resorption of the matrix and replacement with tissue synthesized at least in part by the transfected cells.

Abstract: The invention rates to a hepatitis C virus (HCV) cDNA-based culture system capable of synthesis of infectious HCV in cell culture and cell-to-cell spread of the virus. The invention also relates to a method of measuring the level of HCV infection in a hepatocyte cell. A method for identifying a modulator of HCV activity is also presented, and a method for modulating HCV activity. The invention provides a reliable system for both genetic analysis of the viral genome and for the development of novel antiviral strategies.

Abstract: It has been discovered that there are at least two significant antigens present on the cells of animal species such as pigs that elicit an immune or inflammatory response immediately upon implantation into humans or contact with human serum. The first is an ?-galactosyl (Gal) epitope, for example, Gal?(1->3)Gal?(1->4)GlcNac (linear B type 2) or Gal?(1->3) Gal?(1->4)Glc (linear B type 6). The second is an N-glycolylneuraminic acid (NeuGc) structure. By eliminating these epitopes, preferably by genetically engineering the animal so that the epitope is either not produced or is greatly reduced, or by chemical or enzymatic treatment of the animal's cells to remove the epitopes, it is possible to produce organs, tissues and cells suitable for xenotransplantation into humans.

Abstract: The present invention concerns methods for de-differentiation and stabilize cells, such as progenitor cells, by introducing the cells into the cytoplasm of host oocytes, such as Xenopus laevis oocytes. Advantageously, this method obviates the need for any nuclear transfer procedure, which is known to disrupt the chromosomal architecture of donor and recipient cells. The present invention also concerns host oocytes encapsulating cells that have been introduced into their cytoplasm. The present invention also concerns cells that have been removed from the host oocytes.

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: Methods to obtain genetic modifications of cells in histoculture are described. Modification is assisted by treating the histoculture with collagenase prior to contacting the histoculture with the delivery vehicle for the desired gene. Hair follicles and other organized tissues can be modified in this was and then transplanted into intact recipients.

Abstract: The subject invention is directed to a mixed cell composition to generate a therapeutic protein at a target site by providing a first population of mammalian cells transfected or transduced with a gene that is sought to be expressed, and a second population of mammalian cells that have not been transfected or transduced with the gene, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.

Abstract: A method of expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells by obtaining undifferentiated hemopoietic stem cells or progenitor cells; and either seeding the undifferentiated hemopoietic stem cells or progenitor cells into a stationary phase plug-flow bioreactor in which a three-dimensional stromal cell culture has been pre-established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells, or culturing the undifferentiated hemopoietic stem cells or progenitor cells in conditioned medium obtained from such a reactor.

Abstract: A method of culturing a protein-producing cell, said method comprising co-culturing one transformed cell that can constitutively produce said protein with the parent cell of said transformed cell.

Abstract: A rapid, simple-to-use method for preparing hybrid cells, applicable to fully differentiate, non-dividing cells, entails bringing at least two different cells into contact under conditions that promote cell fusion and then purifying the resultant hybrid without antibiotic or metabolic selection. This approach yields hybrid cells useful in a variety of applications, including clinical treatment regimens, as cellular modulators of the immune system.

Abstract: The present invention relates to a process for activating natural killer cells comprising bringing NK cells into contact with dendritic cells in vitro, ex vivo or in vivo. The invention also relates to cell compositions comprising activated NK cells, NK cell-dendritic cell co-cultures or dendritic cells, and to their use to stimulate the cytolytic activity of NK cells or natural immunity in vivo. The invention also relates to a NK cell stimulation factor present in the dendritic cell membrane, and to triggering media and factor(s) for dendritic cells and to their use, either alone or in combination, to stimulate NK activity, in particular in vivo. The invention can be used to control NK cell activity in vitro, ex vivo or in vivo, in particular under pathological conditions.

Abstract: The present invention relates to a layered cell sorted tissue that is formed in vitro. The tissue is generated by the spontaneous sorting of cells from a homogenous cell mixture into discrete layers by cell type. Connective tissue components, such as fibronectin, may be used to manipulate orientation of the layers during the cell sorting process. The layered cell sorted tissue may be used as a skin graft for burns, wounds, and ulcers. The tissue may also be used in assays to determine effects of chemicals or drugs on human tissue in vitro as well as provide an in vitro assay for tumor cell metastasis.

Abstract: The present invention relates to a biological material having a matrix which contains at least one derivative of hyaluronic acid on which endothelial cells, glandular cells such as islets of Langerhans and liver cells, skin adnexa, germinative cells of hair bulbs, and kerinatocytes are grown, optionally in presence of a medium treated with fibroblasts or in a co-culture with fibroblasts. A process for the production of said biologic materials and the use of such materials for human and veterinary applications such as cardiovascular and oncological surgery, in connection with transplants, for enhancing the biological process of tissue vascularization and for aesthetic use, and also for the screening of medicaments or toxic substances and as a support in the process of gene transfection. The biological material is based on an efficacious cell culture and a biocompatible and biodegradable three-dimensional matrix containing a hyaluronic acid derivative.

Abstract: The present invention provides a model for studying the development of, and/or pathologies associated with neurodegenerative diseases, and agents that can alter such development and/or pathologies. The model of the invention is especially useful as an Alzheimer's disease model. The model of the invention provides brain cells and a method for increasing neurodegenerative disease characteristics in such cells, especially, induction of neurofibrillary tangles and/or phosphorylated tau and/or tau fragments and/or the production and/or release of cytokines and/or microglia reactions and/or activations and/or inflammation and/or conversion of p35 to p25 and/or the levels and activities of protein kinases by selectively increasing the concentration of cathepsin D to an effective level, and/or by lowering the concentration of cholesterol in such cells.

Abstract: According to the present invention, there is provided a biological chamber system having a biochamber defined by outer walls of Sertoli cells. Also provided is a transplantation facilitator including a biochamber. A method of making biochambers by co-culturing facilitator cells and therapeutic cells and then aggregating the facilitator celes is also provided. Also provided is a method of transplanting cells by incorporating transplant cells into a biochamber and transplanting the biochamber containing the transplant cells.

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: Methods and compositions are provided for screening candidate antiviral agents using cells containing subgenomic viral replication systems such as replicons and minigenomes. The methods involve the simultaneous assay of more than one subgenomic viral replication system. Compositions useful for these methods are also provided.

Abstract: The present invention is directed to an apparatus that can be used for co-culturing bacteria and eukaryotic cells. The apparatus allows the bacteria to be grown under steady state conditions and then perfused over the eukaryotic cells. The invention also includes a variety of methods for studying the attachment and invasion of host eukaryotic cells by bacteria.

Abstract: Muscle cells and methods for using the muscle cells are provided. In one embodiment, the invention provides transplantable skeletal muscle cell compositions and their methods of use. In one embodiment, the muscle cells can be transplanted into patients having disorders characterized by insufficient cardiac function, e.g., congestive heart failure, in a subject by administering the skeletal myoblasts to the subject. The muscle cells can be autologous, allogeneic, or xenogeneic to the recipient.

Abstract: Immunostimulatory compositions that contain fused cells formed by fusion between dendritic cells and non-dendritic cells, methods of using these compositions, and methods of generating dendritic cell hybrids.

Abstract: The invention provides cell lines which are useful for the rapid detection and production of influenza and parainfluenza viruses. In particular, the invention relates to transgenic mink lung cells which show increased sensitivity to infection by influenza A, influenza B, or parainfluenza 3 viruses, or which are capable of enhanced productivity of infectious virions. The invention is suitable for use in culturing clinical influenza and parainfluenza virus isolates and for the production of influenza and parainfluenza virus for vaccine formulations, as antigen preparations for diagnostic applications, and for screening antiviral drugs.

Abstract: The present invention provides assays to identify compounds that affect microglial cell activation, and specifically assays to identify compounds that affect secretion of cytokines from these microglial cells by modulating PGE2-mediated activity. The assays of the invention include assays for testing microglial cell activation by contacting microglia with compounds that modulate &bgr;-amyloid PGE2-mediated activation, which can be identified by cellular activity such as secretion of cytokines, e.g., TNF-&agr; and IL-1&agr;. The effect of the candidate compound can be determined by comparing the effect with a control culture which is not contacted with the compound, or by comparing the effect with a standardized profile.

Abstract: A device for performing a biological modification of a fluid, the device includes (a) a chamber having an inlet for intake of the fluid and an outlet for outflow of the fluid; and (b) a collection of micro-organ cultures of at least one organ for performing the biological modification of the fluid, each individual micro-organ culture of the collection including cells and having dimensions, such that cells positioned deepest within the individual micro-organ culture are at least about 100 micrometers and not more than about 225 micrometers away from a nearest surface of the individual micro-organ culture, thereby in vivo organ architecture (organ structure) of organ units (e.g., acinus of liver) is maintained within each individual micro-organ culture, the collection of micro-organ cultures being located within the chamber and the collection of micro-organ cultures being in contact with at least a portion of the fluid flowing through the chamber.

Abstract: The present invention concerns a mammalian cell line which when co-cultured with lymphocytes during which allogenic stimulation is avoided activates lymphocytes fo form tumoricidal cells, a process for the production of tumoricidal T lymphocytes by co-culturing lymphocytes with this cell line, the tumoricidal T lymphocytes obtained by means of this process and the use of the cells according to the present invention for the production of a therapeutic agent which can be used in tumour therapy.

Abstract: A method for inducing differentiation of monocytes contained in an extracorporeal quantity of a subject's blood into functional dendritic antigen presenting cells is provided. The monocytes are first treated by exposure to physical perturbation, irradiation in the presence of a photoactivatable agent capable of forming photoadducts with cellular DNA components, and/or treatment with a DNA binding agent. The treated monocytes are then incubated for a period of time sufficient to maximize the number of functional dendritic cells in the treated cell population. Functional dendritic cells generated from induced monocytes are incubated together with disease effector agents to enhance the presentation of at least one disease-causing antigen expressed by the disease effector agents.

Abstract: By using a bed material for cell culture having a surface composed of two domains of domain A coated with a temperature-responsive polymer and domain B composed of any one or a combination of a domain coated with a polymer having high affinity with cells, a domain coated with the temperature-responsive polymer in an amount different from the amount of the temperature-responsive polymer of domain A, and a domain coated with a polymer which responds to a temperature different from the temperature to which domain A responds, a method for the co-culture of a plurality of kinds of cells which has heretofore been difficult becomes possible.

Abstract: A method for testing the effects of various factors on human skin equivalent is disclosed. The method involves providing a human stratified squamous epithelial cell culture of an immortalized human keratinocyte cell line that forms a reconstructed epidermis, exposing the reconstructed epidermis to a factor and evaluating the effect of the factor on the reconstructed epidermis. A method for selecting preventive or therapeutic agents for skin damages caused by a factor using the same human stratified squamous epithelial cell culture system is also disclosed.

Abstract: Methods of isolating and cryopreserving progenitors from human liver are disclosed which include processing human liver tissue to provide a substantially single cell suspension comprising progenitors and non-progenitors of one or more cell lineages found in human liver; subjecting the suspension to a debulking step, which reduces substantially the number of non-progenitors in the suspension, and which provides a debulked suspension enriched in progenitors exhibiting one or more markers associated with at least one of the one or more cell lineages; and selecting from said debulked suspension those cells, which themselves, their progeny, or more mature forms thereof express one or more markers associated with at least one of the one or more cell lineages. Among these markers are CD14, CD34, CD38, CD 45, and ICAM.

Abstract: A multicellular in vitro assay which models the combined stages of angiogenesis is disclosed. The assay permits the evaluation of drug candidates for their potential utility in treating pathological conditions such as chronic dermal ulcers, tumors, diabetic retinopathy, psoriasis and inflammation.

Abstract: A device for performing a biological modification of a fluid, the device includes (a) a chamber having an inlet for intake of the fluid and an outlet for outflow of the fluid; and (b) a collection of micro-organ cultures of at least one organ for performing the biological modification of the fluid, each individual micro-organ culture of the collection including cells and having dimensions, such that cells positioned deepest within the individual micro-organ culture are at least about 100 micrometers and not more than about 225 micrometers away from a nearest surface of the individual micro-organ culture, thereby in vivo organ architecture (organ structure) of organ units (e.g., acinus of liver) is maintained within each individual micro-organ culture, the collection of micro-organ cultures being located within the chamber and the collection of micro-organ cultures being in contact with at least a portion of the fluid flowing through the chamber.

Abstract: A method for producing human dendritic cells for therapeutic purposes which allows culture-deriving dendritic cells using no cytokines, or reduced cytokines. The method involves culturing mononuclear cells from blood or bone marrow in a medium containing at least one agent such as a calcium ionophore, e.g. A23187, theophylline, protaglandin E1, dibutyryl cyclic AMP, Vitamin D3, Vitamin E, retinoic acid, or a fatty acid. The culture is maintained for a sufficient time, typically 4-14 days, to produce a culture enriched for dendritic cells, as evidenced by at least about 2.5% of total cells exhibiting dendritic cell processes, or a dendritic dell antigen such as CD80, CD86, or CD1a. Also provided is a method to produce antigen-specific human T-cells by pulsing the dendritic cells obtained by the method of the invention with an antigen such as a viral, tumor, bacterial, or cell surface antigen, and then co-culturing T-cells with the antigen-pulsed dendritic cells.

Abstract: A method for testing the effects of various factors on human skin equivalent is disclosed. The method involves providing a human stratified squamous epithelial cell culture of an immortalized human keratinocyte cell line that forms a reconstructed epidermis, exposing the reconstructed epidermis to a factor and evaluating the effect of the factor on the reconstructed epidermis. A method for selecting preventive or therapeutic agents for skin damages caused by a factor using the same human stratified squamous epithelial cell culture system is also disclosed.

Abstract: A device for performing a biological modification of a fluid, particularly in order to assist or replace the functioning of an organ which normally performs this modification, including a collection of liver micro-organ cultures. The device of the present invention is preferably directly connected to a subject for performing this modification.

Abstract: Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. The medium may be conditioned by stromal cells, parenchymal cells, mesenchymal stem cells, liver reserve cells, neural stem cells, pancreatic stem cells and/or embryonic stem cells. Additionally, the cells may be genetically modified. A three-dimensional tissue construct is preferred. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder.

Abstract: This invention provides a method to enhance alloactivation in a mixed lymphocyte culture. Alloactivated cells are effective in treating tumors when implanted into a tumor site or coinjected with tumor cells as a vaccine. By enhancing alloactivation, more cell combinations achieve a threshold of activation adequate for use in therapy, and the level of cytokine secretion in proliferative phase cultures is generally increased.

Abstract: The present invention concerns a mammalian cell line which when co-cultured with lymphocytes during which allogenic stimulation is avoided activates lymphocytes fo form tumoricidal cells, a process for the production of tumoricidal T lymphocytes by co-culturing lymphocytes with this cell line, the tumoricidal T lymphocytes obtained by means of this process and the use of the cells according to the present invention for the production of a therapeutic agent which can be used in tumour therapy.

Abstract: The present invention generally relates to the field of diagnostic microbiology, and, more particularly, to compositions and methods for detecting and differentiating one or more viruses or other intracellular parasites present in a specimen. The present invention also provides compositions and methods to evaluate the susceptibility of a organisms to antimicrobial agents.

Abstract: A method of producing undifferentiated avian cells expressing an embryonic stem cell phenotype. The method includes the steps of collecting avian gonadal cells comprising primordial germ cells from an avian embryo after the formation of the primitive streak; depositing the avian gonadal cells in contact with a preconditioned feeder matrix; and growing the avian gonadal cells on the pre-conditioned feeder matrix in the presence of media for a time sufficient to produce an avian cell culture consisting essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype.

Abstract: The present invention generally relates to the field of diagnostic microbiology, and, more particularly, to compositions and methods for detecting and differentiating one or more viruses or other intracellular parasites present in a specimen. The present invention also provides compositions and methods to evaluate the susceptibility of a organisms to antimicrobial agents.

Abstract: A spontaneously immortalized human kerazinocyte 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 invention relates to cells or tissues having an increased amount of regulatory proteins, including cytokines, growth factors, angiogenic factors and/or stress proteins, and methods of producing and using those cells or tissues. The invention is based on the discovery that the production of regulatory proteins is induced in cells or tissue constructs following cryopreservation and subsequent thawing of the cells or constructs. The compositions and methods of this invention are useful for the treatment of wound healing and the repair and/or regeneration of other tissue defects including those of skin, cartilage, bone, and vascular tissue as well as for enhancing the culture and/or differentiation of cells and tissues in vitro.

Abstract: A novel, empirically derived composition of cytokines and myoblasts is described, that allows for the migration of myoblasts through connective barriers, along with methods employing the composition in the in vivo migration of myoblasts for therapeutic purposes and gene therapy, as well as methods for the identification of agents that are agonistic or antagonistic to myoblast migration in vitro or in vivo.