Abstract: This invention provides methods for preparing novel mammalian multipotent stem cells (MSCs), compositions thereof, and methods of preparing and administering the cells.

Abstract: The present invention is directed to a method of treating cells by co-culturing them with activated fibroblasts in order to regulate the growth and/or status of the cells. Fibroblasts are activated by culturing the cells under conditions that induce the cells to adhere to each other to form multicellular aggregates or spheroids. The present invention also provides a device for selecting cells from cell samples, such as bone marrow aspirate, the device comprises said multicellular aggregates.

Abstract: Methods for wound healing or tissue regeneration by means of cell and tissue engineering, including using three-dimensional matrices with cells therein. A three-dimensional matrix, optionally containing cells such as fibroblasts, is inserted Into the wound of a subject. An anti-inflammatory factor may also be used to reduce or suppress the immune response. The wound may be covered to limit exposure to gaseous oxygen, for example, using a membrane. An anticoagulant may also be applied. In addition, cells, such as fibroblasts or stem cells, when cultured within a three-dimensional matrix, under certain conditions, can be induced to form non-fibroblast multipotent cells. When stem cells are cultured in the three-dimensional matrix, at least some of the stem cells remain as stem cells and do not differentiate. Kits for promoting the control of cells within three-dimensional matrices are also disclosed.

Abstract: This invention relates to novel mammalian multipotent neural stem cells (MNSCs), compositions thereof, and methods of preparing and administering the cells to diseased, aged or damaged tissue such that the cells properly migrate and differentiate and a neurological or corporal deficit is improved or remedied as a result.

Abstract: Disclosed are methods for detecting antibody in a sample, where the antibody targets an antigen expressed by red blood cells or red blood cell ghosts. Rather than detecting the binding events between a particular antigen antibody pair (as in traditional agglutination based assays) the methods herein allow for multiplexed detection of clinically important allo-immune antibodies to blood group antigens. Specifically the method involves generating fluorescently encoded red blood cells or red blood cell ghosts with known antigen presentation and using them to detect the presence of antibody in serum/plasma with a fluorescent sandwich type immunoassay. The assay results can be read using flow cytometric or fluorescent microscope based imaging techniques.

Abstract: An object of the present invention is to develop a method for purify cardiomyocytes at a high degree of purification and at a high yield from a cell mixture comprising cardiomyocytes derived from fetuses and stem cells using various features which have not been previously expected to be used for purification of cardiomyocytes or which are newly found, wherein said method is carried out without undergoing any genetic modification or without adding any special proteins or biologically active agents.

Abstract: Methods for the treatment of cancer with therapies targeting tumor-associated macrophage activities are provided. Methods for the treatment of cancer, inflammatory and autoimmune disorders with therapies using tumor-associated macrophages and adipose tissue macrophages are also provided.

Abstract: Insulin resistance is a central feature of type II diabetes and other diseases, and may affect every tissue of the body, including the pancreatic beta cell. Insulin signaling is mediated by a complex network of diverging and converging pathways, with alternative proteins and isoforms at almost every step in the process. We have previously shown that insulin activates the transcription of its own gene by signaling through Insulin Receptor A type (Ex11?), PI3 kinase and p70 s6 kinase. When studying the mechanisms underlying the glucose-stimulated activation of the glucokinase gene in pancreatic beta cells, we now demonstrate that also here secreted insulin is a key-factor. In contrast to the insulin gene, transcription of the glucokinase gene is promoted by signaling via Insulin Receptor B type (Ex11+) and protein kinase B (c-Akt).

Abstract: This invention relates to the reprogramming of animal cells and animal cell nuclei by nuclear addition. The invention also relates to the generation of animal cells, cell lines, tissues, organs, embryos and non-human animals by methods involving nuclear addition.

Abstract: The invention concerns a method for culturing cells derived from the adipose tissue and in particular the stromal vascular fraction (SVF) to induce formation of cardiomyocytes, the use of the cells obtained by said culture method to reconstitute an ischemized cardiac zone, in particular following an infarction, as well as a pharmaceutical composition containing said cells. The method for obtaining cardiac cells comprises at least the following steps: a) selecting cardiomyogenic cells from the stromal vascular fraction (SVF); b) culturing the cells selected at step a) in a liquid medium optimized for expanding ex vivo the cardiomyogenic cells; c) maintaining and expanding said cells by successive passes in the liquid medium; and d) obtaining cardiac cells.

Abstract: This invention relates to methods of producing a substantially homogenous population of mesenchymal stem cells derived from embryonic stem cells. Also, disclosed is a homogenous population of mesenchymal stem cells capable of further differentiating into a variety of specific cell types, characterized by various morphological factors and cell-specific markers. The compositions and methods described in this disclosure are useful for a variety of commercially important diagnostic, drug screening, and therapeutic applications.

Abstract: The present invention is of methods of establishing and propagating human embryonic stem cell lines using feeder cells-free, xeno-free culture systems and stem cells which are capable of being maintained in an undifferentiated, pluripotent and proliferative state in culture which is free of xeno contaminants and feeder cells.

Abstract: The present invention relates to an oocyte and/or embryo culture medium. The medium includes 0.0003 to 750 ng/ml IGF-II, or a variant or analogue thereof, and further includes either or both of 0.01 to 50 ?g/ml plasminogen, or a variant or analogue thereof, and 0.01 to 50 ?g/ml urokinase plasminogen activator, or a variant or analogue thereof.

Abstract: The disclosure provides methods and compositions to build living tissue covered stents and the like. These tissue coated stents provide a barrier against cell migration to the lumen of the vessel. Since the tissue can surround and envelope the stent, foreign body responses to the stent material are reduced and delayed. The tissue coating is also relatively impermeable to transmural flow, so the wrapped stent can act as a bypass vessel. The tissue is also robust enough to act as a stand alone vessel, without requiring the presence of the metallic stent. These stents can be endothelialized to reduce thrombosis. The genetic modifications described in this disclosure allow for functional organs to be built that express agents that are anti-restenotic or anti-thrombogenic.

Abstract: The present invention is based on the discovery that teeth primordia can be generated using bone marrow cells and that bone marrow cells may be employed to generate teeth without the need for purification and expansion of a population of cells.

Abstract: Induction of neuronal fate in neural stem cells or neural progenitor or precursor cells, or other stem cells, and enhancement of induction of a specific neuronal phenotype, and particularly to induction and enhancement of induction of a midbrain dopaminergic neuronal phenotype. Expressing a nuclear receptor of the Nurr1 subfamily above basal levels within the cell, and regulating GSK-3? inhibition within the cell other than by means of a ligand for a Frizzled receptor.

Abstract: A method for the in vitro production of a cell population P? from a cell population P, the production requiring the presence of at least one factor which is expressed by feeder cells, wherein a) feeder cells proliferate at a temperature T1, b) proliferated feeder cells are contacted with the cell population P, c) the cell mixture obtained at step (b) is cultivated at a temperature T2 which is chosen such that the cell population P proliferates and the feeder cells do not proliferate, the at least one factor being expressed by the feeder cells, and d) the cell population P? so produced is recovered. Advantageously, the production consists in an expansion, the feeder cells are insect feeder cells and the cell population P to be expanded is a T lymphocyte population, preferably a Trl lymphocyte population.

Abstract: A method for producing autonomously contractile heart muscle cells by cultivating and differentiating stem cells obtained from differentiated exocrine gland tissue of an organism is described. Various uses of the heart muscle cells, in particular in regenerative medicine, are also described.

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: The present invention relates to the induction of differentiation in stem cells to cardiomyocytes and factors such as prostaglandin alone or in combination with other factors including essential minerals selected from the group including transferrin and selenium, small molecules selected from the group including a p38 MAPK inhibitor such as SB203580 and protein growth factors of the FGF, IGF and BMP families such as but not limited to IGF1, FGF2, BMP2, BMP4 and BMP6. and insulin that influence the process of differentiation to cardiomyocytes. Media that is appropriate for the induction of differentiation of cardiomyocytes from stem cells is also provided wherein the media contains these factors. The use of cardiomyocytes and cardiac progenitors produced by the directed differentiation in transplantation and screening for cardiac compounds is also provided.

Abstract: To provide a technique of the differentiation from a primate embryonic stem cell into a vascular cell, and techniques using the same. A method for differentiating a primate embryonic stem cell into a vascular cell, comprising differentiating a primate embryonic stem cell into a VEGFR-2-positive and undifferentiated primate embryonic stem cell marker-negative cell, and if need, further differentiating the resulting cell, a method of the differentiation into a vascular cell, and a vascular cell obtained by the method.

Abstract: This invention relates to methods of producing hair folliclesin vitro, compositions for producing hair follicles in vitro, in vitro produced hair follicles, methods of providing an in vitro produced hair shaft at an interfollicular or intrafollicular site, methods of treating hair loss by providing an in vitro produced hair shaft at an interfollicular or intrafollicular site and assays for studying the effect of test agents on hair biology. The invention also provides the similar methods and products which are, or use, immature follicles (“defined herein as proto-hairs”). The invention provides a method for in vitro production of a hair follicle or a proto-hair comprising co-culturing dermal papilla cells with keratinocytes, and optionally with melanocytes.

Abstract: The present invention provides a biocompatible bilayer porous matrix and preparation thereof. The bilayer porous matrix is composed of gelatin, chondroitin 6 sulfate, and hyaluronic acid, also, prepared through freeze-drying technique at different temperature and time duration to form varied pore sizes on each layer. The present invention also provides a method of cell culture using the bilayer porous matrix.

Abstract: The present invention provides novel methods for immunotherapy. The invention provides immune cells and methods to generate them, with the capacity to at least in part reduce an immune response in a host. In one aspect, the invention provides a method for generating a dendritic cell with the capacity to tolerize a T-cell for antigen the T-cell was specific for including culturing peripheral blood monocytes from an individual to differentiate into dendritic cells, activating the dendritic cells in the presence of a glucocorticoid hormone and loading the activated dendritic cell with the antigen the T-cell was specific for.

Abstract: A method of culturing tissue comprises growing tissue forming cells whilst subjecting the tissue forming cells to mechanical stresses which are generated magnetically.

Abstract: The present invention relates to methods for cultivating dermal fibroblasts, methods for preparing in vitro dermis equivalents, methods for preparing three-dimensional in vitro skin equivalents, an in vitro dermis equivalent, a three-dimensional in vitro skin equivalent, and methods for determining the effect of a chemical substance or of an agent on human skin cells using the in vitro dermis equivalent and/or the in vitro skin equivalent.

Abstract: A gene encoding a polypeptide having an activity to support proliferation or survival of hematopoietic stem cells or hematopoietic progenitor cells is isolated by comparing expressed genes between cells which support proliferation or survival of hematopoietic stem cells or hematopoietic progenitor cells and cells which do not support the proliferation or survival. Proliferation or survival of hematopoietic stem cells or hematopoietic progenitor cells is supported by using stromal cells in which the isolated gene is expressed or a gene product of the isolated gene.

Abstract: The invention provides methods for producing a culture of cardiomyocytes and cultures of cardiomyocytes. Exemplary methods of producing and cultures of cardiomyocytes include a population of cells including cells having spontaneous and periodic electrical activity, and/or including nodal, sino-atrial or pacemaker cells; immature cardiomyocytes (cardiomyoblasts); mature contractile cardiomyocytes; or a mixed population of two or more of such cells.

Abstract: The invention relates to methods for inducing marrow stromal cells to differentiate into neural cells by way of increasing intracellular levels of cyclic AMP. The invention also encompasses methods of producing a neural cell by causing a marrow stromal cell to differentiate into a neural cell by increasing intracellular levels of cyclic AMP. Methods for treating a human patient in need of neural cells are also disclosed, as well as methods for treating a human patient having a disease, condition, or disorder of the central nervous system.

Abstract: The present invention provides methods of producing a clone non-human mammalian nuclear transfer (NT) embryo and methods for producing a cloned non-human mammal. Embodiments of the methods include introducing doner genetic material into a metaphase I oocyte; introducing donor genetic material into a non-enucleated oocyte; introducing donor genetic material obtained from a donor cell that is a metaphase into an oocyte; introducing donor genetic material into an oocyte, and naturally activating the oocyte or the NT embryo; and introducing donor genetic material obtained from a donor cell that is at late G1 phase into anoocyte.

Abstract: Compositions and methods for modulating proliferation and/or lineage commitment of stem cells by modulating the Wnt signalling pathways. Modulators of the Wnt signalling pathways and screening methods to identify modulators are also provided. The methods of the invention may be conducted in vitro or in vivo to induce or inhibit stem cell proliferation and/or lineage commitment, and are particularly useful for in vivo stimulation of proliferation and/or lineage commitment of resident stem cells in a tissue.

Abstract: The present invention relates to a renal carcinoma cell line capable of activating the immune system in an antigen-specific manner. According to a further aspect, the invention also includes derivatives of the cell line that maintain said activation capacity. The invention also comprises a method for targeting and activating immune system cells against cells of clear cell renal carcinoma. Said method comprises the co-incubation of isolated immune system cells (dendritic cells, CD4+, CD8+ lymphocytes etc.) with cells of the RCC BA85#21 line in accordance with the invention in a suitable culture medium, for a time sufficient to obtain antigen specific cells.

Abstract: A method of expanding undifferentiated hemopoietic stem cells to increase the number of the hemopoietic stem cells is provided. The method comprising: (a) obtaining the undifferentiated hemopoietic stem cells; and (b) culturing the undifferentiated hemopoietic stem cells in a medium containing a stromal cell conditioned medium, the stromal cell conditioned medium being derived from a stationary phase plug-flow bioreactor in which a three dimensional stromal cell culture has been established under continuous flow of a culture medium 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 the undifferentiated hemopoietic stem cells.

Abstract: A disulfide trap, comprising an antigen peptide covalently attached to an MHC class I heavy chain molecule by a disulfide bond extending between two cysteines, is disclosed. In some configurations, a disulfide trap, such as a disulfide trap single chain trimer (dtSCT), can comprise a single contiguous polypeptide chain. Upon synthesis in a cell, a disulfide trap oxidizes properly in the ER, and can be recognized by T cells. In some configurations, a peptide moiety of a disulfide trap is not displaced by high-affinity competitor peptides, even if the peptide binds the heavy chain relatively weakly. In various configurations, a disulfide trap can be used for vaccination, to elicit CD8 T cells, and in multivalent MHC/peptide reagents for the enumeration and tracking of T cells. Also disclosed are nucleic acids comprising a sequence encoding a disulfide trap. Such nucleic acids, which can be DNA vectors, can be used as vaccines.

Abstract: The invention relates to the method for producing antipeptide cytotoxic T lymphocytes (CTLs) by attaching a complex of a class I HLA and a peptide to antigen presenting cell(s) (APCs) present in a sample of peripheral blood lymphocytes (PBLs), optionally removing excess class I HLA/peptide complex, and incubating with proliferative cytokine. The invention further relates to a CTL obtainable by these methods and to a method of treating a subject by administering such a CTL to the subject.

Abstract: A method of screening for a protein involved in the extracellular regulation of latent TGF? activation by transducing a cell line with a retroviral cDNA library to create a reporter cell line that produces green fluorescent protein (GFP) in response to TGF-? signaling; growing individual clones created by the reporter cell line; co-culturing each individual clone with a second TGF-? reporter cell line that produces luciferase in response to TGF-?, wherein the luciferase production identifies positive clones; and identifying a mechanism of latent TGF-? activation that is employed by the positive clones. A TGF? reporter cell line including a cell line; and a retroviral cDNA library, wherein the reporter cell line produces GFP in response to TGF-? signaling. A method of screening for gene products involved in a biological process.

Abstract: Methods, systems and kits are provided for the clinical staging of blood disorders including myelodysplastic syndrome, myeloproliferative diseases and leukemias by differential analysis of hematologic samples for the distribution of one or more hematopoietic stem or progenitor cell subsets. Additional functional, genetic, gene expression, proteomic or other molecular analyses of the hematopoietic stem and progenitor cells from the patients can also be employed in the staging methods of the invention.

Abstract: Methods for promoting dopaminergic neuronal development and producing neural cells having a dopaminergic phenotype. Dopaminergic neural cells may be used for treating individuals having a neurodegenerative disease such as Parkinson's disease. Dopaminergic cells may be implanted into the brain of the individual, and/or dopaminergic neural development may be induced or enhanced in the brain of the individual. Methods comprise expressing a nuclear receptor of the NG4A subfamily, e.g. Nurr1, above basal levels within the cell and treating the cell with a Wnt ligand, thereby producing or enhancing proliferation, self-renewal, survival and/or dopaminergic induction, differentiation, survival or acquisition of a neuronal dopaminergic phenotype. The cell may be co-cultured with astrocytes or glial cells and may be contacted with an FGF growth factor.

Abstract: Methods for obtaining multipotent mesenchymal stem cells under serum-free conditions and methods for identifying multipotent mesenchymal progenitor cells are disclosed.

Abstract: A method of obtaining a mixture of cells enriched in hepatic progenitors is developed which comprises methods yielding suspensions of a mixture of cell types, and selecting those cells that are classical MHC class I antigen(s) negative and ICAM-1 antigen positive. The weak or dull expression of nonclassical MHC class I antigen(s) can be used for further enrichment of hepatic progenitors. Furthermore, the progenitors can be selected to have a level of side scatter, a measure of granularity or cytoplasmic droplets, that is higher than that in non-parenchymal cells, such as hemopoietic cells, and lower than that in mature parenchymal cells, such as hepatocytes. Furthermore, the progeny of the isolated progenitors can express alpha-fetoprotein and/or albumin and/or CK19. The hepatic progenitors, so isolated, can grow clonally, that is an entire population of progeny can be derived from one cell. The clones of progenitors have a growth pattern in culture of piled-up aggregates or clusters.

Abstract: The invention relates to a method for the non-invasive selection of single living cells under gentle conditions from mixtures of cells or cell cultures with respect to a specific production performance. To this end, the concentration of substances produced by the individual cells which become enriched at the cell membrane, such as reporter gene products (GFP) or specifically secreted products, such as antibodies, is determined by fluorescence-microscopic detection methods.

Abstract: The invention relates to the use of a cultured stem cell to produce a tooth progenitor cell.

Abstract: The present invention is directed to methods which can be used to test agents for their ability 5 to modulate cell proliferation, differentiation and other biological activities of target cells such as stem cells. In addition, the invention is also directed to methods for modulating a biological activity of target cells in vitro, to the cells whose biological activity is modulated by such methods, to the cells that may be produced from target cells by such methods, and to the use of such cells for transplantation, transfusion and other purposes.

Abstract: The present invention relates to methods and uses of cells for the prevention and treatment of a wide variety of diseases and disorders and the repair and regeneration of tissues and organs using low passage and extensively passaged in vitro cultured, self-renewing, colony forming somatic cells (CF-SC). For example, adult bone marrow-derived somatic cells (ABM-SC), or compositions produced by such cells, are useful alone or in combination with other components for treating, for example, cardiovascular, neurological, integumentary, dermatological, periodontal, and immune mediated diseases, disorders, pathologies, and injuries.

Abstract: The present invention is directed to a method of growing new bone, bone-like tissue or extracellular matrix under in vitro cell culture conditions. The method utilizes a bioreactor allowing for the flow of nutrient solutions into, through, and out of the bioreactor, wherein ground demineralized bone and bone-forming cells are present in the bioreactor. The resulting bone, bone-like tissue or extracellular matrix produced by the invention are within the scope of the present invention. In addition, the present invention is directed to the bioreactor device used to grow the new bone, bone-like tissue, or extracellular matrix.

Abstract: A method of interfering with quorum sensing regulation of genes to promote cell growth is disclosed. The method of is aimed at accessing microbial biodiversity. The method involves obtaining an environmental sample comprising at least one novel (uncultivated in the laboratory) microorganism, contacting the environmental sample with an effective amount of an agent or combination of agents which interferes with the quorum sensing regulation of genes, growing the treated sample in a culture medium containing the quorum sensing signal disrupting agent or agents, and analyzing the colonies of microorganisms grown to demonstrate genetic novelty.

Abstract: It is an object of the present invention to provide a feeder cell with less variation in quality. The present invention relates to a feeder cell derived from a tissue stem and/or progenitor cell. A method of preparation of the feeder cell, a method of preparation of a cultured cell using the feeder cell, and a cell culturing kit are also provided.

Abstract: The invention relates to an immortalized feeder cell line. The immortalized feeder cell line may be derived from an embryonic fibroblast, which may be a mouse embryonic fibroblast. A culture of an immortalized feeder cell line according to the invention in a suitable culture medium is also provided, as is a composition including an immortalized feeder cell line according to the invention in a suitable carrier or diluent and conditioned medium produced from growth of an immortalized feeder cell line according to the invention. The invention further provides a method of culturing a stem cell including use of a cell line or conditioned medium according to the invention and cells so produced.

Abstract: Parenchymal cells are cultivated on cultivated endothelial cells or cultivated fibroblasts which have been separated by a surface of a specific hydrophilic polymer, and which have been patterned. A culture which contains thus formed patterned spheroids of cultivated parenchymal cells is thereby provided by this invention. This culture maintains a function which is specific to the parenchymal cells over a long period of time.

Abstract: Parenchymal cells are cultivated on cultivated endothelial cells or cultivated fibroblasts which have been separated by a surface of a specific hydrophilic polymer, and which have been patterned. A culture which contains thus formed patterned spheroids of cultivated parenchymal cells is thereby provided by this invention. This culture maintains a function which is specific to the parenchymal cells over a long period of time.