Abstract: Cyclic amidinium containing compounds and their methods of preparation are described. Compositions containing these compounds facilitate delivery of biologically active polymers to cells in vitro and in vivo.

Abstract: What is described is a method for preparation of insulin-producing cells from non-insulin-producing cells. Mammalian fetal hepatocytes or hepatic progenitor cells are used ad the non-insulin-producing cells, and the method comprises culturing the mammalian fetal hepatocytes or the hepatic progenitor cells with 1-50 mmol/L of nicotinamide and concurrently bringing about expression of the PDX-1 gene or the NeuroD gene in the mammalian fetal hepatocytes.

Abstract: A serum-free C3A clonal cell line and methods for generating the same are provided. The C3A cell line has a reduced doubling time in serum-free medium compared to a corresponding C3A cell line from which it is derived. Methods using the cells of the serum-free C3A clonal cell line for the production, expression and recovery of harvestable polypeptides, screening compounds for metabolic activity, studying enteric disease and for use in a bio-artificial liver device are also provided.

Abstract: Provided are adenoviral vectors and the production of such vectors. In particular, fiber shaft modifications for efficient targeting of adenoviral vectors are provided. The fiber shaft modifications can be combined with other modifications, such as fiber knob and/or penton modifications, to produce fully ablated (detargeted) adenoviral vectors. A scale-up method for the propagation of detargeted adenoviral vectors is also provided.

Abstract: The present invention provides an adenovirus serotype 30 (Ad30) fiber amino acid sequence. The present invention also provides polynucleotides and expression vectors encoding an Ad30 fiber and viral particles and cells containing such expression vectors. The present invention further provides methods of treating genetic diseases or cancers in a mammal using the polynucleotides, polypeptides, expression vectors, viral particles and cells of the present invention.

Abstract: The present invention relates to a new complex receptor polypeptide LSR (Lipolysis Stimulated Receptor), characterized by its functional activities, the cloning of the cDNAs complementary to the messenger RNAs encoding each of the subunits of the multimeric complex, vectors and transformed cells, methods of diagnosis and of selection of compounds which can be used as medicament for the prevention and/or treatment of pathologies and/or of pathogeneses such as obesity and anorexia, hyperlipidemias, atherosclerosis, diabetes, hypertension, and more generally the various pathologies associated with abnormalities in the metabolism of cytokines.

Abstract: The hepatitis C virus (HCV) cell culture system according to the invention consists of human hepatoma cells, which are transfected with a HCV-RNA construct, that comprises the HCV specific RNA segments 5′to NTR, NS3, NS4A, NS4B, NS5A, NS5B, and 3′to NTR as well as a minimum of one marker gene for selection (selection gene).

Abstract: A method of controlling the differentiation of a cell is provided which comprises modulation of the expression of the gene Raidd in the cell. The method provides a means for preparing or enriching a population of stem cells through modulation of Raidd expression.

Abstract: The present invention provides clonal pluripotent hepatic stem cells using flow cytometry and in vitro single-cell-based assays. These cells possess multilineage differentiation potential and self-renewing capability; These cells could be clonally propagated in culture, where they continuously produced hepatocytes and cholanglocytes as descendants while maintaining primitive stem cells. When cells that expanded In vitro were transplanted into recipient animals, they morphologically and functionally differentiated into hepatocytes and cholanglocytes, with reconstitution of hepatocyte and bile duct structures. Furthermore, these cells differentiated into pancreatic ductal and acinar cells or intestinal epithelial cells when transplanted into pancreas or duodenal wall. These data indicate that self-renewing multipotent stem cells persist in the developing mouse liver and that such cells can be Induced to become cells of other organs of endodermal origin under appropriate microenvironment.

Abstract: The present invention provides methods and compositions for supplementing or replacing a damaged organ. The damaged organ to be supplemented or replaced in accordance with the present invention include, for example, kidney, heart, liver, spleen, pancreas, bladder, ureter and urethra. In one embodiment, the tissue-engineered construct of the invention has has at least the following characteristics: (a) differentiated cells on a three-dimensional biocompatible scaffold, wherein the differentiated cells originated from transferred pluripotent cells; and (b) at least one physiological function of the organ.

Abstract: Method for screening for an antiviral agent, by determining whether a potential agent interacts with a virus or cellular component which allows or prevents preferential translation of a virus RNA compared to a host RNA under virus infection conditions; and determining whether any interaction of the agent with the component reduces the level of translation of an RNA of the virus.

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: Methods and compositions that use the hepatitis B virus genome, and fragments or extensions, in a baculovirus vector, to develop anti-HBV agents and to drive high-level expression of a desired gene in a cell of hepatic origin.

Abstract: The invention features modular chambers for culturing cells in which the volume of a chamber can be adjusted without compromising the seal or sterility of the chamber. The invention is based on the principle that the volume of a chamber formed between two plates sandwiching a compressible gasket and a substantially incompressible stop can be adjusted using a gasket that forms a fluid-tight seal between the plates at a plurality of levels of compression. The invention enables the culture of cells between substantially parallel and rigid plates in which a relatively large volume can be used to seed the cells and the holdup volume reduced for perfusion without opening or otherwise disassembling the system to compromise its liquidtightness and sterility. The new closed, modular and scalable cell-culturing chamber can be thus perfused and used to culture cells (e.g., hepatocytes) with high levels of cell function in organ (e.g.

Abstract: The subject invention a method for converting liver stem/progenitor cells to a pancreatic functional cell by transfecting said liver cells with a pancreatic development gene and/or by culturing with pancreatic differentiation factors. The resulting cells produce and secrete insulin protein in response to glucose stimulation.

Abstract: A method of preparing small hepatocytes is provided. The method is suitable for cryopreservation wherein the hepatic function and proliferation ability of the small hepatocytes is retained. A method of cryopreserving thus prepared small hepatocytes and the cryopreserved small hepatocytes having hepatic functions and proliferation ability are also provided. According to the present invention, small hepatocytes are cultured using a medium supplemented with nicotinamide to form colonies of small hepatocytes in which the small hepatocytes are encompassed by nonparenchymal cells, and then the formed colonies are dissociated from culture dishes as small hepatocytes aggregate by non-enzymatic treatment, suspended in a cryopreservation solution and are cryopreserved.

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.

Abstract: A cell support system useful for the implantation of living cells in a subject comprises a solid substrate, typically formed from a biologically inert material. The substrate has a textured surface portion, with the textured surface portion defining a plurality of recessed cavities therein. A plurality of live cells to be implanted are deposited on the textured surface portion so that the cells (or progeny thereof) are protected from mechanical dislodgment.

Abstract: The invention provides methods, compositions and devices for inducing tissue-specific regeneration in a mammal, or for stimulating proliferation of mammalian progenitor cells. The present methods, compositions and devices make use of osteogenic protein 1 (OP-1), which is appreciated herein as a tissue morphogen, i.e., a substance competent to induce tissue-specific morphogenesis of mammalian body tissues in addition to bone and/or cartilage. Alternatively, the present methods, compositions and devices make use of other naturally-occurring or biosynthetic proteins sharing a defined structural and functional relationship with OP-1 and thus appreciated herein also to function as tissue morphogens. Optionally, OP-1 or a related protein can be used alone or when adsorbed on a support matrix which provides an anchoring substratum for proliferation and/or differentiation of progenitor cells during tissue-specific morphogenesis.

Abstract: A surgical procedure (partial hepatectomy) in the nonhuman primate allows repeated removal of large liver tissue samples from a single animal for the preparation of liver tissue slices and primary cell culture of hepatocytes. The open biopsy procedure, alternating non-anatomic and anatomic partial liver resection monthly on the same liver lobe, follows a surgical protocol allowing animal welfare, survival, and multiple sampling (at least 8) from the same animal, thus reducing considerably the number of animals needed for research purposes.

Abstract: The invention relates to new flow-through cell culturing devices that include plates arranged in parallel and spaced to control the flow patterns and velocity of liquid medium flowing between the plates. The devices can also include gas-permeable, liquid impermeable membranes arranged between the plates. The devices can be used to culture cells, such as hepatocytes, at high levels of mass transport of nutrients, oxygen, and waste products, yet low levels of shear stress for extended periods of time and with high levels of cell function, e.g., in organ assist systems. The invention also includes new culturing plates for use in the devices, and methods of manufacturing these plates.

Abstract: Disclosed are methods for isolating dendritic cells and/or dendritic progenitor cells. The methods include contacting a population of cells with a plurality of FRIL family member molecules, and removing the unbound cells, wherein the cells bound to the FRIL family member molecules are an isolated population of dendritic cells and/or dendritic progenitor cells.

Abstract: This invention relates to methods of isolating hepatoblasts utilizing panning techniques and fluorescence activated cell sorting. This invention further relates to isolated hepatoblasts and to a method of treating liver dysfunction as well as to methods of forming artificial livers.

Abstract: A defective recombinant adenovirus including at least one DNA sequence coding for all or an active part of a superoxide dismutase or a derivative thereof. The therapeutical use thereof and corresponding pharmaceutical compositions are also disclosed.

Abstract: Methods and associated materials for transducing mesenchymal stem cells with a desired nucleic acid. Mesenchymal stem cells are a recently discovered kind of stem cell for which suitable transfer vehicles are still desired. Typical gene delivery vehicles such as the adenoviruses or adeno associated viruses have no particular tropism for mesenchymal stem cells. Also disclosed is gene therapy using adenoviruses provided with tropism for mesenchymal stem cells.

Abstract: This invention provides a system for producing differentiated cells from a stem cell population for use wherever a relatively homogenous cell population is desirable. The cells contain an effector gene under control of a transcriptional control element (such as the TERT promoter) that causes the gene to be expressed in relatively undifferentiated cells in the population. Expression of the effector gene results in depletion of undifferentiated cells, or expression of a marker that can be used to remove them later. Suitable effector sequences encode a toxin, a protein that induces apoptosis, a cell-surface antigen, or an enzyme (such as thymidine kinase) that converts a prodrug into a substance that is lethal to the cell. The differentiated cell populations produced according to this disclosure are suitable for use in tissue regeneration, and non-therapeutic applications such as drug screening.

Abstract: A method of propagating mammalian endodermally derived progenitors such as hepatic progenitors, their progeny, or mixtures thereof is developed which includes culturing mammalian progenitors, their progeny, or mixtures thereof on a layer of embryonic mammalian feeder cells in a culture medium. The culture medium can be supplemented with one or more hormones and other growth agents. These hormones and other growth agents can include insulin, dexamethasone, transferrin, nicotinamide, serum albumin, &bgr;-mercaptoethanol, free fatty acid, glutamine, CUSO4, and H2SeO3. The culture medium can also include antibiotics. Importantly, the culture medium does not include serum.

Abstract: A substantially enriched mammalian hepatic liver engrafting cell population is provided. Methods are provided for the isolation and culture of this liver engrafting cell. The progenitor cells are obtained from a variety of sources, including fetal and adult tissues. The cells are useful in transplantation, for experimental evaluation, and as a source of lineage and cell specific products, including mRNA species useful in identifying genes specifically expressed in these cells, and as targets for the discovery of factors or molecules that can affect them.

Abstract: Treatment of stem cells with a retinoid induces differentiation of the stem cells into hepaticopancreatic tissue.

Abstract: The present invention relates to a method for increasing survival rate of cells in animal cell culture under hypoxia condition by adding antibiotics to the culture media. The method of present invention comprises a step of culturing animal cells in culture media containing antibacterial agent of quinolones, quinones, aminoglycosides or chloramphenicol at the concentration range of 0.1 to 1000 &mgr;g/ml. The invented method can be practically applied for high-density animal cell culture to produce recombinant proteins or cultured cells.

Abstract: Novel fetal genes (fls353 and fls485) have been successfully isolated from human fetal liver-derived cDNAs. These genes were specifically expressed in tissues including fetal tissues which are thought to contain a large number of undifferentiated cells and actively differentiating/proliferating cells. High levels of expression of these genes were observed also in a variety of cancer cells. The proteins and genes encoding the proteins can be used as the tool for developing drugs for the treatment of tumors.

Abstract: The invention relates to the discovery of a novel RNA sequence at the 3′ terminal sequence of hepatitis C virus (HCV) genome RNA. Included in the invention are the 3′ sequence, its complement, and their use for nucleic-acid based diagnostics and for developing and evaluating novel anti-HCV therapies. This sequence element, which is conserved among HCV genotypes, is likely to be essential for viral replication, and required for construction of full-length HCV cDNA clones capable of yielding infectious RNA, progeny virus or replication-competent HCV replicons. Such functional clones are useful tools for evaluation of therapeutic approaches and as substrates for developing candidate attenuated or inactivated HCV derivatives for vaccination against HCV.

Abstract: The invention features-modular cell culturing devices including one or more flat-plate modules, and is based on the discovery that if the flows of liquid medium and oxygenated fluid are separated by a gas-permeable, liquid-impermeable membrane, and the cells are grown attached to the liquid side of the membrane, the device can be used to culture cells with transport of oxygen through the membrane (i.e., direct oxygenation), without regard for the flow rate of the liquid medium passing through the device. The new flow-through cell culturing devices can thus be used to culture cells, e.g., hepatocytes, with high levels of cell function in organ, e.g., liver, assist systems, for production of cells, for production of cell-derived products, such as, proteins or viruses, or for systems to treat biological liquids to remove toxins, such as, ammonia, or add cell-synthesized products, or both.

Abstract: A method for culturing Langerhans islets to obtain an amount sufficient for transplant and autotransplant is disclosed. The islets are cultured in a culture serum (rat/human) medium which is supplemented with radical scavengers, growth factors, a matrix material, nerve growth factor, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at proper the time during the culturing process. The medium is supplemented with radical scavengers and growth factors for the first time and then further supplemented with matrix material, radical scavengers, nerve growth factor and the growth factors around 12-24 hours after culturing. Thereafter, the medium is supplemented with growth factors, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at 4-5 days into the culturing process. The culturing process is conducted for an extended period of time, so that any latent red blood cells are eliminated from the islet culture.

Abstract: Antisense oligonucleotides that hybridize to segments of the pres1, S, C, and &egr; regions of the hepatitis B virus (HBV) RNA pregenome inhibit replication of the virus. Pharmaceutical compositions which contain these oligonucleotides as the active ingredients are effective against HBV infection.

Abstract: It has been discovered that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. In one example, human embryonic stem cells are allowed to form embryoid bodies, and then combined with the differentiation agent n-butyrate, optionally supplemented with maturation factors. In another example, n-butyrate is added to human embryonic stem cells in feeder-free culture. Either way, a remarkably uniform cell population is obtained, which is predominated by cells with morphological features of hepatocytes, expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function.

Abstract: The present invention relates to a biological material comprising a matrix consisting of at least one derivative of hyaluronic acid on which endothelial cells, glandular cells such as islets of Langerhans and liver cells, skin adnexa, germinative cells of hair bulbs, and keratinocytes are grown, optionally in presence of a medium treated with fibroblasts or in a co-culture with fibroblasts. It is also described the process for the production of said biologic material and its use for human and veterinary use, in cardiovascolar and oncological surgery, in transplants, to enhance the biological process of tissue vascularization and for aesthetic use, and also for the screening of medicaments or toxic substances and as a support gene transfection.

Abstract: The invention provides a method of treating liver damage or disease in a patient by stimulating liver regeneration. Specifically, the invention provides a method of inducing liver cell proliferation comprising contacting liver cells that express FoxM1B protein with growth hormone. The invention also provides methods of screening for compounds that induce FoxM1B protein expression, nuclear localization, or both expression and nuclear localization. The invention further provides pharmaceutical compositions comprising selected compounds and methods of using such compositions.

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

Abstract: It has been discovered that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. In one example, human embryonic stem cells are allowed to form embryoid bodies, and then combined with the differentiation agent n-butyrate, optionally supplemented with maturation factors. In another example, n-butyrate is added to human embryonic stem cells in feeder-free culture. Either way, a remarkably uniform cell population is obtained, which is predominated by cells with morphological features of hepatocytes, expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function.

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 compositions comprising cells that can effectively produce HCV after HCV infection, compositions for culturing the cells, methods for making the composition and methods for infecting the cells in the composition with HCV. The present invention also provides methods for assaying HCV production and methods for evaluating compounds that affect the production of HCV.

Abstract: This application discloses methods and materials for preparing functional microvascular beds in the laboratory. These prevascularized constructs can be used to vascularize engineered tissue constructs or to revascularize damaged or diseased tissues or organs following implantation. The prevascularized constructs may also deliver genetically engineered gene products to the bloodstream.

Abstract: This application discloses methods and materials for preparing functional microvascular beds in the laboratory. These prevascularized constructs can be used to vascularize engineered tissue constructs or to revascularize damaged or diseased tissues or organs following implantation. The prevascularized constructs may also deliver genetically engineered gene products to the bloodstream.

Abstract: It has been discovered that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. In one example, human embryonic stem cells are allowed to form embryoid bodies, and then combined with the differentiation agent n-butyrate, optionally supplemented with maturation factors. In another example, n-butyrate is added to human embryonic stem cells in feeder-free culture. Either way, a remarkably uniform cell population is obtained, which is predominated by cells with morphological features of hepatocytes, expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function.

Abstract: The present invention provides three novel HSC genes designated SCM 3, SCM 26, and SCM 113, the coding regions thereof, the gene products, applications of the genes, DNA constructs, vectors and transformed cells each comprising the gene of a fragment thereof. Methods of using the SCM 3, SCM 26 and SCM 113 polynucleotide and polypeptide sequences are also disclosed.

Abstract: A non-naturally occurring method of modulating the targeting a pluripotent stem cell to a target tissue of a mammalian subject from another site in the mammalian subject includes the step of: increasing or decreasing the concentration of SDF-1 alpha protein in the target tissue.

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: Hepatocyte cells that are reversibly immortalized and grown in culture, which are functional and safe for use in transplantation, are disclosed. Also disclosed are methods for immortalizing primary hepatocytes, expanding the population of immortalized hepatocytes in culture, then reversing the immortalization to produce hepatocytes that are functional and safe for use in transplantation.