Abstract: The present invention relates to a pigment epithelial cell of the eye containing vector DNA of an adenoviral vector with large DNA capacity, to the improved isolation and cultivation of these cells and to methods for production and the use in the therapy of an eye or nerve disease.

Abstract: Artificial dermis (1) obtained from plasma with platelets (2) and human fibroblasts. The plasma with platelets (2) is obtained from the fractionating of total blood (4) from the patient (8) by light centrifugation, and the human fibroblasts (3) from a skin biopsy (5). Clotting is obtained by adding calcium. This artificial dermis (1) provides for the rapid growth of the keratinocytes (6) seeded on its surface to build an artificial skin (7) which can easily be transplanted. Large areas of artificial dermis (1) are obtained from a small skin biopsy (5) and minimal quantities of plasma with platelets (2), which being enriched with cytokines and platelet growth factors, strengthens the proliferation of the cells seeded, both inside and on the surface. The artificial skin (7) obtained can be used to treat major burn treatments, chronic skin ulcers, etc., or be used, by employing genetically altered cells, as a vehicle for gene therapy.

Abstract: Disclosed is a method of preparing an undifferentiated cell. The method includes contacting a more committed cell with an agent that causes the more committed cell to retrodifferentiate into an undifferentiated cell.

Abstract: An organotypic culture comprises an artificial stroma overlayed with epithelial cells isolated from a human colon or intestine. The stroma comprises a mixture of collagen and human fibroblasts isolated from a human colon or intestine. The culture contains a factor that binds the IGF-1 receptor, a factor that binds the EGF receptor, and a factor that binds the LIF receptor. These factors may be added exogenously to the culture via medium or may be expressed by various recombinantly engineered cell types in the culture. The organotypic culture can result in growth that is in situ-like or emphasizes other physiological or morphological states, depending on the balance of factors in the growth media. The organotypic culture may be used in methods for screening of therapeutic, carcinogenic, or growth enhancement factors, or for treating intestinal injuries by applying to the site of an injury the intact culture or the components thereof.

Abstract: The invention relates to a method for lymphoid tissue-specific cell production from hematopoietic progenitor cells in unique, three-dimensional culture devices, in the presence of antigen presenting cells and lymphoreticular stromal cells, and in the absence of exogenously added growth factors. The resulting lymphoid tissue-specific cells may be isolated at any sequential stage of differentiation and further expanded. The lymphoid tissue-specific cells also may be genetically altered at any stage of the process.

Abstract: This invention provides a method for rapidly culturing a large amount of human chondrocytes to give normal chondrocytes or a mass thereof. The culture method comprises co-culturing human chondrocytes together with perichondral cells in the chondrogenic stage, as feeder cells, which support the proliferation ability of the chondrocytes, to allow rapid culturing of the human chondrocytes in a large amount.

Abstract: The present invention provides methods for culturing normal and malignant human bladder epithelial cells for many generations, and compositions of matter to be used in the methods.

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: This invention provides medicaments and methods for managing cancer using donor cells that are alloactivated in culture. Alloactivated cells are implanted into the bed of a solid tumor and initiate a response by the host against the tumor. Subsequently, alloactivated cells are implanted into the bed of a solid tumor a second time. The two implants work synergistically to confer remarkable benefit to the treated subject, both in terms of management of the cancer and the development of an anti-cancer immune response. The beneficial effects may include regression of the tumor and extended survival. Removal of any residual tumor after the second implant facilitates ongoing resistance to tumor regrowth or metastasis.

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 methods and compositions useful for delivering antigens to dendritic cells which are then useful for inducing antigen-specific cytotoxic T lymphocytes and T helper cells. This invention also provides assays for evaluating the activity of cytotoxic T lymphocytes. According to the invention, antigens are targeted to dendritic cells by apoptotic cells which may also be modified to express non-native antigens for presentation to the dendritic cells. The dendritic cells which are primed by the apoptotic cells are capable of processing and presenting the processed antigen and inducing cytotoxic T lymphocyte activity or may also be used in vaccine therapies.

Abstract: A method for providing a urinary tract tissue graft composition includes providing a tissue culture frame and a segment of small intestinal submucosa and positioning the segment of small intestinal submucosa in the tissue culture frame such that the segment of small intestinal submucosa is suspended and held in a taut position by the tissue culture frame. Smooth muscle and urothelial cells are isolated from a tissue specimen of a subject and cultured, and then seeded upon the segment of small intestinal submucosa, thereby forming a urinary tract tissue graft. A tissue culture frame in which such a urinary tract tissue graft may be formed is also disclosed.

Abstract: Cell compositions consisting essentially of mammalian hematopoietic CXCR4+ stem and progenitor capable to migrate in response to stromal-derived factor 1 (SDF-1) and/or capable to adhere to stromal cells in response to an adhesion-inducing agent, are provided for clinical transplantation. Hematopoietic CXCR4?/low stem and progenitor cells can be converted into CXCR4+ cells by stimulation with a suitable agent. The composition consists preferably of human CD38?/low CXCR44 cells.

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: A method for constructing a stable bioactive mammalian embryonic kidney is described herein. A kidney so constructed requires no artificial support, nor porous man made membranes or tubing to effectuate its biological function of filtering body fluids. A single donor embryonic kidney, or fragment thereof, can produce a great number of functional kidneys suitable for treating subjects with various kidney disorders. It is anticipated that said in vitro produced kidney would be less, or not at all, antigenic when transplanted into a subject, because of its embryonic character and artificial propagation in culture. This method of producing a functional organ can be useful in cloning other organ structures containing inducible epithelial tissues.

Abstract: The present invention provides an antigen-presenting cell(APC)/tumor cell conjugate, wherein the antigen-presenting cell (APC) is modified by a cytokine gene selected from the group consisting of IL-2, IL-3, IL-4, IL-6, IL-12, IL-18, IFN?, IFN?, IFN?, TNF, TGF, GM-CSE, and the combination thereof. The conjugate is useful as a tumor vaccine to significantly induce an immunity specifically against the tumor cell The present invention also provides the method for preparing the conjugate and a pharmaceutical composition containing said conjugate.

Abstract: The present invention relates to the development of insulin resistant skeletal muscle cell culture model useful for the screening of compounds that enhance insulin receptor (IR) and insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation required against type II diabetes.

Abstract: A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (?); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation are human embryonic stem cells, have normal karyotypes, and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.

Abstract: A method of producing a homogenous population of homozygous stem (HS) cells pre-selected for immunotype and/or genotype from donor cells is described herein. The invention relates to methods of using immunohistocompatible HS cells for diagnosis, therapeutic and cosmetic transplantation, and the treatment of various genetic diseases, neurodegenerative diseases, traumatic injuries and cancer. The invention further relates to methods for using histocompatible HS stem cells pre-selected for a non-disease genotype for prophylactic and therapeutic intervention including, but not limited to, therapeutic and cosmetic transplantation, and the treatment of various genetic diseases, neurodegenerative diseases, and cancer.

Abstract: In this application is described the establishment and maintanence of a normal human hepatocyte cell line able to support complete development of malaria parasite development in vitro. Advantages and uses of the cell line are also described.

Abstract: Biological tissues are grown in a low shear, microgravity environment by culturing connective tissue cells to form a three-dimensional structure, which is thereafter co-cultured with endothelial and epithelial cells to replicate naturally occurring tissues. Preferably, the three-dimensional connective tissue cells are first cultured with endothelial cells to form three-dimensional structures of connective tissue cells and endothelial cells, which are thereafter co-cultured with epithelial cells to replicate naturally occurring tissue. The cultured tissue is in the general shape of spheroids having a diameter between about 0.1 mm and about 5 m.

Abstract: The invention relates to the field of medicine and more particularly it relates to the problem of vaccination against tumor cells and vaccinotherapy of oncological diseases, and also to a method of treating diabetes mellitus. In the invention a new method of cultivating cells is proposed, which contemplates forming a capsule of a polyacrylamide gel in the tissue of an animal, including a human, into which capsule desirable cells are injected. The invention provides for maintaining the viability of cells during a long period of time.

Abstract: The present invention provides a method for determining expression levels of one or a multiplicity of target proteins in a tissue or cell sample.

Abstract: Novel cells and molecules involved in tumor immunity are disclosed. The novel cells are regulatory T-cells having the phenotype CD3+??-TcR+CD4?CD8?CD44?CD28?NK1.1?. The regulatory cells express Ly6A and osteopontin while non-regulatory cells do not.

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: Disclosed is a cervico-vaginal tissue equivalent comprised of vaginal epithelial cells and immune cells, cultured at the air-liquid interface. The tissue equivalent is capable of being infected with a sexually transmitted pathogen such as a virus (e.g., HIV), a bacteria, a helminthic parasite, or a fungus. The tissue equivalent is also capable of undergoing an allergic-type reaction or an irritant-type reaction. The tissue equivalent is characterized as having nucleated basal layer cells and nucleated suprabasal layer cells, and further as having cell layers external to the suprabasal layer progressively increasing in glycogen content and progressively decreasing in nuclei content. Immune cells of the tissue equivalent are primarily located in the basal and suprabasal layers. Also disclosed are methods for producing the tissue equivalent.

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 cell or tissue-culturing carrier which can effectively regenerate an intended a cell or tissue, while suppressing an excessive growth of fibroblasts, and a method of culturing a cell or tissue by using the above carrier, the cell or tissue-culturing carrier wherein fibroblasts showing substantially no growing property in a gel based on the hydrogel-forming polymer, is constituted by using a hydrogel-forming polymer; an aqueous solution of which shows a thermo-reversible sol-gel transition such that it assumes a sol state at a lower temperature and assumes a gel state at a higher temperature.

Abstract: The present invention relates to the use of therapeutic compounds in the modification of T-cells, T-cell-antigen presenting cell (APC) interactions and the interaction between pathogenic organism and immunocompetent cells of a host. In particular, it relates to the use of these compounds in the modulation of the interaction between Notch proteins and their ligands and to the use of such compounds in the therapy of conditions such as graft rejection, autoimmunity, allergy, asthma and infectious diseases.

Abstract: The present invention relates to a method of in vitro spermatogenesis involving Sertoli cells and diploid germ cells from a testis of a male mammal to yield differentiated haploid spermatids. The present invention also relates to spermatids produced by the method described above, where the spermatids are haploid. The present invention also involves a method of overcoming male infertility in mammals involving the use of the haploid round spermatids produced by the in vitro spermatogenesis method of the present invention. The present invention also relates to isolated haploid spermatids.

Abstract: Nitric oxide adversely affects survival and development of cells such as oocytes and embryos in vitro, particularly in a co-culture system. The addition of a nitric oxide inhibitor such as hemoglobin to such systems eliminates this toxic effect, and promotes mammalian oocytes, embryos, or other cells in vitro.

Abstract: Diagnostic methods for the detection of multiple myeloma (MM) and the identification of high-risk patients with multiple myeloma-related plasma proliferative disorders, such as MGUS or SMM, likely to progress to active MM are described. The diagnosis is based on the determination of concentrations of bioactive IL-1? produced by the bone marrow plasma cells of these patients. Also described are therapeutic methods for the treatment of MM and for the chemoprevention of the progression from disorders such as MGUS and SMM to active MM, involving the administration of inhibitors of IL-1?.

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 in vitro cultured skin substitutes, and in particular to improved methods for organotypic culture of skin substitutes. In some embodiments, the dermal equivalent of the skin substitute is lifted to air interface of the culture prior to seeding with keratinocytes. In other embodiments, increased concentrations of collagen are used to form the dermal equivalent. In still other embodiments, optimized media are utilized to maintain the skin equivalents.

Abstract: This invention relates to methods of using human dendritic cells to present antigens for the induction of antigen-specific T cell-mediated immune responses. In particular, it relates to the isolation of dendritic cells from human blood, exposing the cells to antigens, co-culturing the antigen-pulsed dendritic cells with &ggr;&dgr;-T cell receptor-positive-T cells (&ggr;&dgr;-TCR+ T cells) obtained from unprimed or weakly primed individuals for the stimulation of antigen-specific T cell proliferative and cytotoxic activities. The dendritic cell antigen presentation system described herein has a wide range of applications, including but not limited to, activation and expansion of large numbers of antigen-specific major histocompatibility complex-unrestricted T cells for use in adoptive cellular immunotherapy against infectious diseases and cancer.

Abstract: The present invention relates to novel substrates, to methods of making them and to uses therefor. The substrates of the invention comprise a base portion and a surface layer covering at least part of the base portion, with a binding layer provided therebetween. The surface layer provides, on at least a part of the substrate, topographical features having at least one nano scale dimension. These topographical features are adapted to facilitate, influence, control or inhibit cell or tissue growth thereon and/or therebetween and the substrate may be used for the study, manipulation or modification of at least one cellular or tissue behaviour or response.

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: The present invention relates to a programmable scaffold, which is a three-dimensional scaffold having interconnected pores and biologically active molecules physically entrapped therein. The scaffold is a lyophilized hydrogel of a polymer. The scaffold can be used in an array on a platform and loaded with various combinations of biologically active molecules for high throughput and parallel screening, as well as tissue engineering. The present invention also relates to methods for making and modifying the scaffolds.

Abstract: Methods and apparatus for recovering dental pulp from dentition of a donor are disclosed, wherein the pulp from within extracted teeth utilizing such methods and apparatus is harvested, while preserving a sterile environment and avoiding trauma and infection, and stem cells, dendritic cells, and other cells isolating from the pulp, and the various cells propagated and expanded for subsequent use in repair or regeneration of tissues of the body, for therapeutic treatments, and other medical purposes.

Abstract: A preparation and a method of making composite blastocysts (CBs) from aggregates of dissociated cells of non-viable pre-embryos are disclosed. The CB is characterized morphologically by having two distinct tissue types, the inner cell mass (ICM) and the trophectoderm (TE), and a blastocoelic cavity (BC). The method of making CBs is an aggregation process (AP) comprising inter alia the following steps: 1) dissociation of discarded pre-embryos; 2) isolation of single nucleated cells from dissociated discarded pre-embryos; 3) microsurgical encapsulation of several cells within a host zona pellucida or artificial aggregation with or without a non-zona vessel; and 5) primary culture of the cell aggregates for multiplication and differentiation of cells. One particularly advantageous embodiment is that the starting material is non-viable pre-embryos. Another advantageous embodiment is that the AP allows individual cells from non-viable pre-embryos to further multiply, and become integrated into CBs.

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

Abstract: The differential expression of marker proteins in a targeted population provides a means of identifying and isolating cells. A population of cells associated with the regeneration of pancreatic islets is shown to express certain proteins, including the cell surface proteins ErbB2, ErbB3, and ErbB4; and the nuclear protein Msx-2. Populations of isolated pancreatic islet progenitor cells find use in screening assays, to characterize genes involved in islet development and regulation, and in transplantation to provide a recipient with pancreatic islet functions.

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 culture chamber of the present invention has a fluid-filled culture compartment in which cells, tissues and other biologicals are cultured. The culture compartment is transversed by one or more molecular weight cut-off membranes attached to a membrane carrier assembly. Incoming nutrients are transported through the membrane into the culture compartment and metabolic waste products are transported away from the fluid-filled culture compartment through the membrane and out the chamber outlet. Both reusable and disposable culture chambers are described for culturing cells, cell aggregates, particles, tissues and organoids.

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

Abstract: The invention relates to a cell composition containing macrophages, presenting anti-infectious and hematopoietic properties. More particularly, the invention relates to a cell composition containing macrophages, myeloid cells and progenitors; said cell compositions are useful for the restoration of hematopoiesis in an aplasic patient and/or the protection of patients against infectious diseases or against residual tumors.

Abstract: Human bone marrow-derived stem cells are differentiated into pancreatic endocrine marker-expressing cells in vitro by first culturing human bone marrow mononuclear cells in a tissue culture container to obtain cells that adhere to the container, and continuing to culture the adherent cells until they become morphologically homogenous; second, culturing the morphologically homogeneous cells in a medium containing high glucose levels at least until the cells express detectable levels of glucagon, insulin, and mRNAs encoding insulin, Pdx-1, and NeuroD; and third, culturing the cells in a medium containing low glucose levels, nicotinamide, and exendin 4. Transplantation of pancreatic endocrine marker-expressing cells made in this manner can reduce hyperglycemia in a diabetic animal.

Abstract: A method of repairing tissue of an organ in a patient's body includes co-culturing pluripotential stem cells obtained from the patient's body with cells obtained from a site other than the patient's body (i.e., non body cells) and having specific functions of the tissue to be repaired for mimicking by the stem cells and creating a specific microenvironment, maintaining the culture for a period of time sufficient for modification of the stem cells by acquisition of the specific functions of the non body cells; segregating the modified stem cells from the non body cells, and implanting only the modified stem cells to and at a site of the tissue to be repaired.

Abstract: The present invention relates to a medium for preparing dedifferentiated cells derived from post-natal islets of Langerhans. The medium comprises in a physiologically acceptable culture medium an effective amount of a solid matrix environment for a three-dimensional culture, a soluble matrix protein, and a first and a second factor for developing, maintaining and expanding the dedifferentiated cells. Such a medium may be used in an in vitro method for islet cell expansion.