Abstract: The present invention relates to cell adhesion promoting (“CAP”) peptide combinations that promote cell attachment or cell adhesion to culture surfaces that are otherwise cell adhesion resistant “CAR”. The invention provides combination of peptides that, when covalently coupled to a CAR layer such as hyaluronic acid that has been created on a polystyrene surface, promote cell attachment, growth differentiation, and execution of other desired cellular functions in culture.

Abstract: The present invention relates to an assay system and method for testing compounds for their ability to regulate the hepatic lipase (HL) promoter. In particular, the invention relates to the identification of estrogen receptor ligands having this activity. Compounds that inhibit HL promoter activity are useful as leads, or on their own, to develop therapeutics in the prevention of heart disease.

Abstract: The present invention relates to methods of screening drug candidates for toxic effects on human cells. The invention provides methods for determining idiosyncratic toxicity of test compounds.

Abstract: The present invention provides a method comprising a step of transferring a cell proliferation factor gene into a mammalian liver cell to obtain an immortalized liver cell, a step of proliferating the immortalized liver cell, and a step of removing the cell proliferation factor gene from the immortalized liver cell; a large number of liver cells obtained thereby; and a treating agent and a artificial liver comprising obtained liver cell.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

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: Replication competent adenoviral vectors specific for cells expressing alfa-fetoprotein (AFP) are provided. These replication-competetent adenoviral vectors comprise adenovirus genes essential for replication under the transcriptional control of an AFP-transcriptional regulatory element.

Abstract: Described are compositions with tethered growth effector molecules, and methods of using these compositions for growing cells and tissues. Growth effector molecules, including growth factors and extracellular matrix molecules, are flexibly tethered to a solid substrate. The compositions can be used either in vitro or in vivo to grow cells and tissues. By tethering the growth factors, they will not diffuse away from the desired location. By making the attachment flexible, the growth effector molecules can more naturally bind to cell surface receptors. A significant feature of these compositions and methods is that they enhance the biological response to the growth factors. The method also offers other advantages over the traditional methods, in which growth factors are delivered in soluble form: (1) the growth factor is localized to a desired target cell population; (2) significantly less growth factor is needed to exert a biologic response.

Abstract: The present invention relates to the preparation of non-human animals having chimeric livers, whereby some or substantially all of the hepatocytes present are human hepatocytes. It is based, at least in part, on the discovery that rats, tolerized in utero against human hepatocytes, were found to serve as long-term hosts for human hepatocytes introduced post-natally, and the introduced hepatocytes maintained their differentiated phenotype, as evidenced by continued production of human albumin. The present invention further relates to the use of such animals as models of various liver diseases, including viral invention. Such embodiments are based on the discovery that transplanted human hepatocytes in chimeric livers were found to be susceptible to Hepatitis B virus and Hepatitis C virus infection.

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 relates generally to an assay for detecting variant Hepatitis B viruses (HBVs) which exhibit altered sensitivity to agents. The variant HBVs are delivered to cells using a baculovirus vector. The altered sensitivity to an agent is in relation to the effects of the agent on one or more stages of infection, replication, assembly or release of virus or virus-like particles. The identification of variant HBVs with altered sensitivities to anti-HBV agents provides a means of monitoring cross resistance, or the development of new therapeutics effective against variant HBVs with altered sensitivities to other anti-HBV agents, as well as monitoring therapeutic protocols. The present invention further provides variant HBVs detected by the assay of the present invention and to components thereof as well as recombinant, chemical analogue, homologue and derivative forms of such components.

Abstract: The present invention relates to the production of islet cells and insulin in a subject by providing for expression of an islet transcription factors in the pancreas of the subject, by for example, introduction of nucleic acid encoding the transcription factor neurogenin3 or a factor that induces neuorgenin3 expression. The present invention also relates to methods for using a islet transcription factor gene and the islet transcription factor polypeptide to alter cellular differentiation in culture or in vivo to produce new ?-cells to treat patients with diabetes mellitus.

Abstract: A unique HCV RNA molecule is provided having an enhanced efficiency of establishing cell culture replication. Novel adaptive mutations have been identified within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture. This self-replicating polynucleotide molecule contains, contrary to all previous reports, a 5?-NTR that can be either an A as an alternative to the G already disclosed and therefore provides an alternative to existing systems comprising a self-replicating HCV RNA molecule. The G->A mutation gives rise to HCV RNA molecules that, in conjunction with mutations in the HCV non-structural region, such as the G(2042)C/R mutations, possess greater efficiency of transduction and/or replication. These RNA molecules when transfected in a cell line are useful for evaluating potential inhibitors of HCV replication.

Abstract: This invention provides a method for determining susceptibility for an anti-viral drug comprising: (a) introducing a resistance test vector comprising a patient-derived segment and an indicator gene into a host cell; (b) culturing the host cell from (a); (c) measuring expression of the indicator gene in a target host cell; and (d) comparing the expression of the indicator gene from (c) with the expression of the indicator gene measured when steps (a)-(c) are carried out in the absence of the anti-viral drug, wherein a test concentration of the anti-viral drug is present at steps (a)-(c); at steps (b)-(c); or at step (c).

Abstract: The present invention relates to a new tumor suppressor, designated CAR-1, the gene for which is located on the short arm of human chromosome 1. This gene is directly implicated in colon, kidney and breast cancers, and the CAR-1 ubiquitous expression of the corresponding transcript suggests that it may be involved in yet others. Thus, one aspect of the invention is the diagnosis of CAR-1-related malignancies. The full length cDNA for CAR-1, as well as oligonucleotides derived therefrom, are disclosed. Screening methods for modulators of CAR-1 function and expression, as well as methods for cancer therapy, are described.

Abstract: AAV vectors may have utility for gene therapy but heretofore a significant obstacle has been the inability to generate sufficient quantities of such recombinant vectors in amounts that would be clinically useful for human gene therapy application. Stable AAV packaging cell lines have been elusive, mainly due to the activities of Rep protein, which down-regulates its own expression and can negatively affect the host cell. This invention provides packaging systems and processes for packaging AAV vectors that effectively circumvent these problems and that allow for substantially increased packaging efficiency.

Abstract: The invention provides an isolated and purified DNA molecule comprising at least one DNA segment, a biologically active subunit or variant thereof, of a circular intermediate of adeno-associated virus, which DNA segment confers increased episomal stability, persistence or abundance of the isolated DNA molecule in a host cell. The invention also provides a composition comprising at least two adeno-associated virus vectors.

Abstract: Artificial liver devices and methods for using the devices to purify a biological fluid are disclosed. The methods include the use of living hepatocytes (23) which are either unattached or attached to inert carriers and suspended in a cell culture medium which circulates in the devices with the hepatocytes (23). Blood or plasma passes on one side (7?) of semi-permeable membranes, on the other side (7) of which is the cell culture medium and across which is a concentration and/or pressure gradient. Solutes diffusing across the membrane into the cell culture medium are metabolized by the hepatocytes (23) and/or captured by additional removal means (4). Those undesirable substances which do not diffuse out of the blood or plasma into the hepatocyte containing culture medium are captured by additional removal means (50).

Abstract: A bioartificial liver system has a separator for separating plasma and blood, a liver-slice culture apparatus, or bioreactor, to detoxify the plasma. The bioreactor has a chamber with plasma and gas inlets, at least two meshes mounted parallel one above the another, near the upper portion of the chamber forming at least two horizontal layers separated by a space. A plurality of liver slices positioned within the space, a supply of plasma is provided to the chamber so that it rises to contact the liver slices, and is alternatively removed from contacting the liver slices, and a supply of gas is provided to the top of the chamber. The system also includes a reservoir for containing plasma entering and exiting the chamber. Methods are provided for detoxifying plasma using the bioartificial liver system.

Abstract: This invention relates to the generation of liver cells and in particular to the repopulation of the liver with healthy cells for patients with liver disease, especially acute liver failure. The invention thus provides a human liver cell progenitor characterised by the following markers: (a) CD 117 positive, and (b) CD 133 positive, and (c) CD 34 negative.

Abstract: Provided are methods of inhibiting proliferation and/or trans-differentiation of hepatic stellate cells, which method comprises the step of contacting hepatic stellate cells with a ligand of the low affinity glucocorticoid binding site (LAGS), which may be the rat p28 receptor or the human hpr6.6 receptor. The invention also provides ligands for use in such methods (e.g. pregnenolone 16-alpha carbonitrile) and methods for screening for the same (e.g. based on competition or displacement assays using LAGS ligands such as dexamethasone). Such ligands may be used in the treatment of liver disorders.

Abstract: An objective of the present invention is to provide a process for effectively producing hematopoietic stem cells. A process for producing hematopoietic stem cells comprises the step of culturing hematopoietic stem cells in the presence of a gp130 stimulating factor, one or more cytokines and stromal cells.

Abstract: A culture medium, which is capable of sustaining long-term cultures of hepatocytes and liver cells. In this medium, mammalian primary hepatocytes retain highly replicative capacity and hepatic gene expression activity. The liver cells from genetically defined sources may be reproducibly immortalized without the delivery of foreign genes, such as viral oncogenes. The immortalized hepatocytes are non-tumorigenic, making them suitable for clinical and therapeutic purposes.

Abstract: The invention features methods of promoting dedifferentiation of pancreatic cells, methods of obtaining pancreatic islet cells from the dedifferentiated pancreatic cells, and methods of treating a subject having a disorder characterized by insufficient pancreatic islet function by administering pancreatic islet cells obtained by these methods.

Abstract: A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.

Abstract: A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.

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: The present invention relates to methods of culturing stem cells to produce hepatocyte-like cells. Among other advances, the invention also relates to purified preparations of hepatocyte-like cells and methods for using the hepatocyte-like cells.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

Abstract: The present invention provides a cell line which can be substituted for &bgr; cells in human mature pancreatic islets and express insulin in a glucose-concentration dependent manner, and enables the easy obtainment of the number of cells which meets the demand. The present invention also provides a therapeutical cell preparation for treating diabetes. The cell lines of the present invention can be obtained by integrating both a nucleotide sequence encoding tamoxifen-induced Cre recombinase and a nucleotide sequence encoding insulin regulated by glucose-sensitive promoter into the chromosome in a human immortalized hepatic cell line FERM BP-7498 containing the TERT gene inserted in between a pair of LoxP sequences.

Abstract: A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.

Abstract: It was discovered that a membrane protein dlk (delta-like) is specifically expressed on the surfaces of fetal hepatic cells. We succeeded in purifying undifferentiated hepatic cells including hepatic stem cells to high purify from mouse fetal liver using an antibody to dlk protein. The obtained fetal hepatic cells included hepatic stem cells having bipotency. The present invention provides a marker molecule of hepatic cells, and a method for detecting undifferentiated hepatic cells utilizing this molecule. The present invention also provides a method for separating undifferentiated hepatic cells utilizing this marker molecule, and the undifferentiated hepatic cells separated by this method. By using the method of the present invention, hepatic stem cells may be purified simply to high concentration. The undifferentiated hepatic cells obtained by the present invention may be used for regenerative medicine and artificial liver.

Abstract: This invention relates to cell lines that are obtained using cultured cell lines derived from human liver as a host and that stably express a number of human cytochromes P450. The human liver-derived cultured cell lines of the present invention are useful in, for example, analyzing an enzyme participating in the metabolism of xenobiotics or endogenous substrates, because of their stable expression of human cytochromes P450 CYP1A1, 1A2, 2A6, 2B6, 2C8, 2C9, 2C9, 2D6, and 3A4.

Abstract: The present invention is directed toward a method for obtaining from whole liver or a resection thereof a population of cells comprising viable, functional liver cells enriched in hepatocytes and hepatocyte stem/progenitor cells, compositions thereof, and uses therefore. Compositions include a composition of liver cells enriched in hepatocytes and hepatocyte stem/progenitor cells and a pharmaceutical composition thereof. Uses include treatment of liver diseases, regeneration of liver, toxicity testing, and liver assist devices.

Abstract: Methods, including culture media conditions, which provide for in vitro human stem cell division and/or the optimization of human hematopoietic progenitor cell cultures and/or increasing the metabolism or GM-CSF secretion or IL-6 secretion of human stromal cells and/or a method for assaying the effect of a substance or condition on a human hematopoietic cell population, and/or depleting the malignant cell or T-cell and B-cell content of a human hematopoietic cell population are disclosed. The methods rely on culturing human stem cells and/or human hematopoietic progenitor cells and/or human stromal cells in a liquid culture medium which is replaced, preferably perfused, either continuously or periodically, at a rate of 1 ml of medium per ml of culture per about 24 to about 48 hour period, and removing metabolic products and replenishing depleted nutrients while maintaining the culture under physiologically acceptable conditions. Optionally, growth factors are added to the culture medium.

Abstract: The present invention is directed to methods for the expansion of non-terminally differentiated cells (“precursor cells”) using agonists of Notch function, by inhibiting the differentiation of the cells without inhibiting proliferation (mitotic activity) such that an expanded population of non-terminally differentiated cells is obtained. The cells are preferably stem or progenitor cells. These expanded cells can be used in cell replacement therapy to provide desired cell populations and help in the regeneration of diseased and/or injured tissues. The expanded cell populations can also be made recombinant and used for gene therapy, or can be used to supply functions associated with a particular precursor cell or its progeny cell.

Abstract: A unique HCV RNA molecule is provided having an enhanced efficiency of establishing cell culture replication. Novel adaptive mutations have been identified within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture. This self-replicating polynucleotide molecule contains, contrary to all previous reports, a 5′-NTR that can be either an A as an alternative to the G already disclosed and therefore provides an alternative to existing systems comprising a self-replicating HCV RNA molecule. The G→A mutation gives rise to HCV RNA molecules that, in conjunction with mutations in the HCV non-structural region, such as the G(2042)C/R mutations, possess greater efficiency of transduction and/or replication. These RNA molecules when transfected in a cell line are useful for evaluating potential inhibitors of HCV replication.

Abstract: The present invention provides a mammalian immortalized liver cell obtained by transferring a cell proliferation factor gene located between a pair of site-specific recombination sequences into a mammalian liver cell.

Abstract: A composition which comprises an animal cell population which contains immature animal cells. The immature animal cells are characterized by expression of alpha-fetoprotein or lack of essential expression of alpha-fetoprotein and albumin, and at least a portion of said immature animal cells or at least a portion of the progeny of said immature animal cells is capable of differentiating into cells which express albumin. The cell population is cultured under conditions which result in expansion of the cells. Expansion of the cells may be achieved by culturing the cells in the presence of an extracellular matrix and liver stromal cells; and preferably in the presence of growth factors. Such cells may be used for liver transplantation, artificial livers, and for toxicology and pharmacology studies. Such cells may also be genetically engineered to express proteins or polypepetides of interest.

Abstract: The present invention provides a method comprising a step of transferring a cell proliferation factor gene into a mammalian liver cell to obtain an immortalized liver cell, a step of proliferating the immortalized liver cell, and a step of removing the cell proliferation factor gene from the immortalized liver cell; a large number of liver cells obtained thereby; and a treating agent and a artificial liver comprising obtained liver cell.

Abstract: The present invention provides a Hepatitis C Virus (HCV) replicon that efficiently replicates in an eukaryotic cell. The HCV replicon includes a nucleic acid sequence encoding a subgenomic fragments of HCV of any genotype that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding an acetyl transferase selectable marker, such as puromycin. Also provided is an HCV type 1a replicon that efficiently replicates in an eukaryotic cell and includes a nucleic acid sequence encoding subgenomic fragments of type 1a HCV that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding a acetyl transferase selectable marker. Further provided are eukaryotic cell lines that include an HCV replicon or an HCV type 1a replicon which efficiently replicate in the eukaryotic cell. The present invention also provides screening methods for identifying candidate compounds that inhibit the propagation of HCV.

Abstract: Hepatic progenitors comprise two populations of human hepatic stem cells, primitive and proximal hepatic stem cells, and two populations of committed progenitors, one for biliary cells and one for hepatocytes. Human primitive hepatic stem cells are a very small fraction of the liver cell population and give rise to proximal hepatic stem cells constituting a much larger fraction of the liver. Human proximal hepatic stem cells give rise to biliary and hepatocyte committed progenitors. Primitive and proximal stem cells are the primary stem cells for the human liver. Human primitive hepatic stem cells may be isolated by immunoselection from human livers or culturing human liver cells under conditions which select for a human primitive hepatic stem cell. Proximal hepatic stem cells may be isolated by immunoselection, or by culturing human liver cells under conditions which include a developmental factor.

Abstract: The present invention is directed to methods for readily generating hepatocyte precursor cell lines that retain hepatocyte-specific functions after extensive in vitro culturing. The methods comprise isolating and culturing hepatocyte precursor cell lines under permissive culture conditions that suppress asymmetric cell kinetics and allow exponential growth of the precursor cells, followed by transferring the hepatocyte precursor cell lines to non-permissive culture conditions that allow expression of asymmetric cell kinetics and induce expression of hepatocyte-specific characteristics.

Abstract: The invention provides cells and methods of using the cells for the propagation of replication-deficient adenoviral vectors. The cells comprise at least one heterologous nucleic acid sequence which upon expression produces at least one non-adenoviral gene product that complements in trans for a deficiency in at least one essential gene function of one or more regions of an adenoviral genome so as to propagate a replication-deficient adenoviral vector comprising an adenoviral genome deficient in the at least one essential gene function of the one or more regions when present in the cell.

Abstract: The present invention relates to recombinant hepatitis C virus (HCV)-derived nucleic acids and to stable cell clones derived from human hepatoma Huh-7 cell line supporting high titer replication of said recombinant HCV nucleic acids. The subgenomic HCV replicons and cell clones of the instant invention represent the in vitro system of choice for studies of HCV propagation, anti-viral drug screening, and vaccine development.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

Abstract: The present invention features novel placental derived stem cells and provides methods and compositions for the therapeutic uses of placental derived stem cells or placental derived stem cells that have been induced to differentiate into a desired tissue type into a recipient host in amounts sufficient to result in production of the desired cell type, i.e., hepatic, pancreatic, neuronal, or nervous tissue.

Abstract: A vascularised tissue construct seeded with endothelial cells is disclosed. A modular, scalable method of fabrication of tissue constructs with a uniform cell distribution that can accommodate multiple cell types, and in which the porosity is created after cell incorporation, is disclosed. The construct is based on the porous structure that is created when a column or tube is randomly packed with solid objects, such as cylindrical rods. Cells, such as liver cells, are encapsulated in solid gelatin rods (200 &mgr;m diameter, aspect ratio 1:5) on to which if desired endothelial cells, such as human umbilical vein endothelial cells (HUVEC), can adhere. The gelatin rods are randomly packed into a larger tube and then coated with HUVEC in one embodiment. The interstitial gaps among the rods form interconnected channels that are lined by the endothelial cells. The resulting endothelial cell lining enables whole blood to percolate around the rods and through the interstitial channels.

Abstract: This invention provides methods of generating cells that stably replicate sub-genomic virus replicons. This invention also provides methods of generating cells that have disabled PKR activity and that stably replicate HCV sub-genomic replicons. The invention also provides methods of using the cells of the invention to screen for compounds that modulate viral RNA replication, including HCV RNA replication.

Abstract: Highly purified hepatic stem cells are trans-differentiated into pancreatic endocrine hormone-producing cells by culturing them in vitro in a medium containing high levels of glucose. These trans-differentiated cells express insulin, glucagon, and pancreatic polypeptide, but not hepatocyte protein Hep-par. When stimulated with glucose, these cells synthesize and secrete insulin, a response enhanced by nicotinamide. Transplantation of these trans-differentiated cells into a hyperglycemic animal normalizes blood sugar levels in the animal.