Abstract: The method of tissue transplantation involving recellularizing of a donor organ with the utilization of the recipient's cytokines collected from the recipient's blood plasma at less than systemic pressure, and at the temperature greater than freezing and less than normal systemic temperature of the recipient's blood. Specifically, a method of harvesting a platelet-derived growth factors from platelet rich plasma (PRP), the growth factors having increased weight.

Abstract: An isolated antibody that has a specific binding affinity to a polypeptide comprising the amino acid sequence HTEGKP (SEQ ID NO: 2) phosphorylated at threonine is provided. The antibody may be used as biomarker for mitotic cells. A method for using the antibody in accordance with the invention comprises contacting a cell with the antibody and detecting antibody bound to the cell as an indicator of the cell being in the mitotic state. A reagent kit comprising the antibody is also provided.

Abstract: Methods and devices for culturing human pluripotent stem cells to produce cells of the pancreatic lineage are disclosed. The methods include steps of culturing the stem cells under conditions that induce the expression of mesendoderm/primitive streak and definitive endoderm markers in a chemically defined medium including an effective amount of i) fibroblast growth factor, ii) Activin A, and iii) bone morphogenetic protein. The methods further include the steps of culturing cells under conditions favoring the formation of at least one of intact embryoid bodies and pancreatic progenitor PDX1+Ins? cells.

Abstract: The present invention relates to synthetic green tea derived polyphenolic compounds, their modes of syntheses, and their use in inhibiting proteasomal activity and in treating cancers. The present invention is also directed to pharmaceutical compositions useful in methods of inhibiting proteasomes and of treating cancers.

Abstract: The present invention generally relates to the modulation of hypoxia-inducible factor (HIF) using the compounds and methods disclosed herein. These compounds and methods can be applied to the prevention, pretreatment, and/or treatment of conditions or states associated with HIF, such as hypoxia- and ischemia-related conditions and the induction of stasis.

Abstract: The present invention provides methods of enhancing hybridoma fusion efficiencies through cell synchronization of the fusion partners, in order to aid in production of antibodies.

Abstract: The present invention provides methods for inducing division of postmitotic mammalian differentiated cardiomyocytes. The invention can be used to repair heart tissue damaged by, for example, myocardial ischemia, hypoxia, stroke, myocardial infarction or chronic ischemic heart disease in vivo. In addition, the methods of the invention can be used to induce heart muscle cells to divide in vitro, in vivo and/or ex vivo, which can then be used in heart tissue repair.

Abstract: The present invention provides methods for regulating the growth and/or survival of tumor cells and stem cells by modulating the expression or function of ATF5. The present invention also provides methods for promoting or suppressing differentiation of stem/progenitor cells, for producing differentiated cells and for isolating/purifying differentiated cells, including neural cells. Also provided are differentiated cells, cell populations and transgenic animals comprising same and uses of same. The present invention further provides methods for treating nervous tissue degeneration and for identifying an agent for use in treating nervous tissue degeneration. Methods for promoting apoptosis in neoplastic cells and for treating or preventing tumors, and identifying agents for use in treating or preventing tumors are also provided by the present invention. The present invention further provides methods for identifying agents that inhibit ATF5, agents identified by these methods.

Abstract: A method of differentiating embryonic stem cells into ventral spinal progenitor cells is disclosed. In one embodiment, the invention comprises culturing a population of cells comprising a majority of cells that are characterized by an early rosette morphology and are Sox1?/Pax6+ in the presence of retinoic acid, wherein the cells express Hoxb4, but not Otx2 or Bf1.

Abstract: Disclosed are methods and pharmaceutical compositions for inducing pancreatic hormone production.

Abstract: This invention relates to methods for distinguishing and sorting cells. In particular it includes methods for distinguishing and sorting post-mitotic and post-meiotic daughter cells into two classes according to differential cellular features. Labeling, tagging, or marking of the cells' chromatin proteins, RNA, or DNA may assist in distinguishing the daughter cells. In some embodiments, two cell classes may be studied and the cells' proteins, glycoproteins, and RNA may be identified and subset. Information from these subsets may then be used to distinguish and sort the two classes of cells from similar tissues according to protein, glycoprotein, and RNA makeup.

Abstract: A system combining a clonogenic differentiation assay with an instrument-based ATP bioluminescence proliferation assay to produce a standardized colony-forming stem and progenitor cell potency assay is provided.

Abstract: A method of preparing neural precursor cells by exposing pluripotent stem cells or neural stem cells to a differentiation agent. The agent is a pyridine analog, which in preferred embodiments is a phenylethynyl-substituted or phenylazo-substituted pyridine. In other embodiments, a method of enhancing neural precursor cell survival is provided in which the survival is enhanced by exposure to the pyridine analog. In further embodiments, a method of preparing neuronal cells is provided in which pluripotent or neural stem cells exposed to the pyridine analog are then incubated without the pyridine analog, resulting in differentiation into neurons, astrocytes and oligodendrocytes. These methods may be used in toxicological screens, e.g., to evaluate the neurotoxicity of a test compound.

Abstract: The present invention relates generally to kits that provide reagent mixes and instructions for the use thereof, in performing high-throughput assay methods that determine the proliferative status of isolated target cell populations. The methods measure the luminescent output derived from the intracellular ATP content of incubated target cells, and correlate the luminescence with the proliferative status of the cells. The present invention further relates to kits that provide reagent mixes and instructions for high-throughput assays methods for screening compounds that may modulate the proliferative status of a target cell population. The kits of the present invention and methods therein described may be used for determining the proliferative status of any isolated cell line or type. The kits and methods of the present invention address the need for rapid assays that determine the proliferative status of isolated hematopoietic stem and progenitor cells and of subpopulations of differentiated cells thereof.

Abstract: The present invention relates to a method of inhibiting differentiation of a population of neural stem cells by contacting a purinergic receptor agonist and a population of neural stem cells under conditions effective to inhibit differentiation of the population of neural stem cells. Another aspect of the present invention relates to a method of producing neurons and/or glial cells from a population of neural stem cells by culturing a population of neural stem cells with a purinergic receptor antagonist under conditions effective to cause the neural stem cells to differentiate into neurons and/or glial cells. The purinergic receptor agonist can also be used in a method of inducing proliferation and self-renewal of neural stem cells in a subject and a method of treating a neurological disease or neurodegenerative condition in a subject. The purinergic receptor antagonist can also be used in treating a neoplastic disease of the brain or spinal cord in a subject.

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

Abstract: A use of a composition comprising umbilical cord blood-derived mesenchymal stem cells for inducing differentiation and proliferation of neural precursor cells or neural stem cells to neural cells is provided, the composition being effective for the treatment of nerve injury diseases.

Abstract: This application relates to a newly identified animal cell structure, the midbody scar. This structure is a remnant of the midbody that is retained by one daughter cell following cytokinesis and persists through multiple subsequent cell cycles. The midbody scar can be useful as a marker of dividing cells or of a cell's replicative age.

Abstract: The present invention relates generally to assays, methods, and kits that provide reagent mixes and instructions for determining the proliferative status of isolated target cell populations. The methods measure the luminescent output derived from the intracellular ATP content of incubated target cells, and correlate the luminescence with the proliferative status of the cells.

Abstract: The invention relates to an antisense oligonucleotide targeted to the coding region of the human acetylcholinesterase (AChE), which selectively suppresses the AChE-R isoform of the enzyme. The antisense oligonucleotide is intended for use in the treatment and/or prevention of neuromuscular disorders, preferably myasthenia gravis. In addition, it can penetrate the blood-brain barrier (BBB) and destroy AChE-R within central nervous system neurons, while also serving as a carrier to transport molecules across the BBB.

Abstract: The present invention relates generally to high-throughput assay methods that determine the proliferative status of hematopoietic stem and progenitor cells. The present invention further relates to high-throughput assays for screening compounds that modulate the growth of hematopoietic stem and progenitor cells and for identifying subpopulations thereof that are suitable for transplantation. The assay of the present invention is particularly useful for quality control and monitoring of the growth potential in the stem cell transplant setting and would provide improved control over the reconstitution phase of transplanted cells.

Abstract: The present invention relates generally to kits that provide reagent mixes and instructions for the use thereof, in performing high-throughput assay methods that determine the proliferative status of isolated target cell populations. The methods measure the luminescent output derived from the intracellular ATP content of incubated target cells, and correlate the luminescence with the proliferative status of the cells. The present invention further relates to kits that provide reagent mixes and instructions for high-throughput assays methods for screening compounds that may modulate the proliferative status of a target cell population. The kits of the present invention and methods therein described may be used for determining the proliferative status of any isolated cell line or type. The kits and methods of the present invention address the need for rapid assays that determine the proliferative status of isolated hematopoietic stem and progenitor cells and of subpopulations of differentiated cells thereof.

Abstract: Methods for collecting cells in M phase or G1 phase, by which the percentage of M phase or G1 phase cells is higher than that attained by the conventional methods are 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: The present invention is a recombinant vector encoding and expressing at least three or more costimulatory molecules. The recombinant vector may additionally contain a gene encoding one or more target antigens or immunological epitope thereof. The synergistic effect of them costimulatory molecules on the enhanced activation of T cells is demonstrated. The degree of T-cell activation using recombinant vectors containing genes encoding three costimulatory molecules was far greater than the sum of recombinant vector constructs containing one costimulatory molecule and greater that the use of two costimulatory molecules. Results employing the triple costimulatory vectors were most dramatic under conditions of either low levels of first signal or low stimulator to T-cell ratios. This phenomenon was observed with both isolated CD4+and CD8+T cells.

Abstract: Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

Abstract: The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Abstract: This invention provides a method of generating antigen specific allospecific human suppressor CD8+CD28− T cells. This invention also provides a method of generating xenospecific human suppressor CD8+CD28− T cells. This invention further provides a method of generating allopeptide antigen specific human suppressor CD8+CD28− T cells. Methods of treatment for reduction of risk of rejection of allografts and xenografts and autoimmune diseases using the human suppressor CD8+CD28− T cells so produced are also provides, as are methods of preventing rejection and autoimmune diseases, and vaccines comprising the produced suppressor T cells. Methods of diagnosis to determine whether a level of immuno-suppressant therapy requires a reduction are provided.

Abstract: The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Abstract: A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.

Abstract: We teach a strategy to obtain large quantities of desired APCs, activated B cells, which are superior in their capacity to present tumor protein antigen in a multiadministration protocol. Human B cells can be obtained from peripheral blood in large numbers. These cells can be activated in vitro by coculture with CD40L (CD40-B cells) and an immunosuppressive agent such as cyclosporin A. They can expanded up to 1×103 to 1×104 fold in 2 weeks or 1×105 to 1×106 fold in 2 months. We demonstrate these cells are most efficient APCs comparable to DCs in stimulating allogeneic CD4+ CD45RA+, CD4+ CD45RO+, and CD8+ T cells. In contrast to DCs, CD40-B cells are fully functional even in the presence of immunosuppressive cytokines such as IL-10 and TGF&bgr;.

Abstract: The present invention relates generally to methods for stimulating cells, and more particularly, to a novel method to concentrate and stimulate cells that maximizes stimulation and/or proliferation of such cells. In the various embodiments, cells are stimulated and concentrated with a surface yielding enhanced proliferation, cell signal transduction, and/or cell surface moiety aggregation. In certain aspects methods for stimulating a population of cells such as T-cells, by simultaneous concentration and cell surface moiety ligation are provided by contacting the population of cells with a surface, that has attached thereto one or more agents that ligate a cell surface moiety and applying a force that predominantly drives cell concentration and cell surface moiety ligation, thereby inducing cell stimulation, cell surface moiety aggregation, and/or receptor signaling enhancement.

Abstract: The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Abstract: A first substance (such as the peptide aldehydes LLnL and LVP) inhibits the function of intracellular proteasome. A second substance (such as ranpirnase) inhibits intracellular protein synthesis or increases intracellular expression of a cyclin-dependent kinase (CDK) inhibitor. Both substances are administered in such a manner that the effects thereof are coincident. The cell cycle of affected tumor cells is therefore arrested and the cells die of apoptosis.

Abstract: Described herein are methods for obtaining from an initial population of cells a plurality of subpopulations of cells having a similar density by growing samples from the initial population in medium having a limiting nutrient. Also provided are methods for using such subpopulation of cells to identify members of an initial population, either spontaneously occurring or induced by mutagenesis, having altered and/or preferred phenotypes.

Abstract: A method is disclosed for gene transfer into a culture of primitive stem cells which comprises a prestimulation step of adding a blocking agent to block at least one inhibitor of a cell cycle of said primitive stem cells. The prestimulation is time-limited for a period of less than approximately 36 hours so that said culture of primitive stem cells retains hematopoietic potential.

Abstract: The present invention relates to a method of synchronizing epithelial cells into G.sub.0 phase, more specifically, a method of synchronizing epithelial cells into G.sub.0 phase characterized by culturing the cells in a synthetic medium for 5 days or more, wherein said synthetic medium is a medium with the lowered concentration of bovine fetal serum 5% or less and said epithelial cells is bovine mammary gland cells. Further, the present invention relates to a method of culturing cells characterized by using G.sub.0 phase synchronized cells obtained by the present invention, and a method of screening a related to differentiation and/or maturation of said cells by using G.sub.0 phase synchronized cells. The present invention is useful for easily synchronizing epithelial cells into G.sub.0 phase and for screening a factor related to differentiation and/or maturation of epithelial cells.

Abstract: Transgenic mice which constitutively express an antibody-type molecule encoded by the transgene and which has an IgE heavy chain constant region and is specific for a pre-defined antigen, provide an allergic reaction to that antigen without prior sensitization and are useful as allergy models.

Abstract: The present invention is directed to an in vitro method for detecting the presence of carcinogenic agents.

Abstract: The present invention is directed to novel chimeric proliferation receptor proteins and DNA sequences encoding these proteins where the chimeric proteins are characterized in three general categories. In one category, the novel chimeric proteins comprise at least three domains, namely, an extracellular inducer-responsive clustering domain capable of binding an extracellular inducer that transmits a signal to a proliferation signaling domain, a transmembrane domain and a proliferation signaling domain that signals a host cell to divide. In the second category, the novel chimeric proteins comprise at least two domains, namely, an intracellular inducer-responsive clustering domain capable of binding an intracellular inducer and a proliferation signaling domain that signals the cell to divide.

Abstract: A system is provided for selective clonogenic expansion of relatively undifferentiated cells, including (a) a tube containing a plurality of beads of a size which permits a plurality of the undifferentiated cells to grow thereon, the beads bearing on their surfaces a selective binding molecule which binds to a surface antigen present on the relatively undifferentiated cells, wherein the antigen is not present on the surfaces of the relatively differentiated cells; (b) means for continuously providing nutrients to the relatively undifferentiated cells growing on the beads, wherein the nutrients are delivered via a fluid which flows through the tube and past the beads; and (c) means for continuously harvesting the relatively undifferentiated cells downstream of the beads.