Abstract: Differentiation of human pluripotent stem cells, such as human embryonic stem cells (hESC), into hepatocytes by in vitro methods is disclosed. The pluripotent stem cells are cultured in conditioned medium from the hepatocarcinoma cell line, HepG2. Specific growth factors and defined media may also be added to the medium for stage specific differentiation of the derived hepatocytes. Hepatocytes differentiated from human pluripotent stem cells may be characterized by fluorescence activated cell sorting (FACS), immunofluorescence analysis (IF), real time polymerase reaction (RT-PCR), and functional assays. The methods disclosed herein are able to differentiate high percentages of hepatocytes from human pluripotent stem cells using the disclosed methods. These differentiated cells may exhibit polygonal shape morphology, typical of hepatocytes, and may express hepatocyte specific genes. The differentiated cells may also be positive for definitive endoderm markers and hepatic markers.

Abstract: Human pluripotent stem cells which are isolated from cut human umbilical cord or placenta and characteristic of cell surface marker CD151+, OCT4+ and CD184?, can adhere to tissue culture plastic and have the potential to differentiate into three germ layers: endoderm, mesoderm and ectoderm. Methods of isolating, purifying and culturally expanding of a population of human pluripotent stem cells and uses for treating diseases caused by cell damage or cell aging, and as a kind of carrier cells in gene therapy are provided.

Abstract: The present invention relates to a method of growing a plurality of cells (110) to performance competence in a matrix forming agent (120) containing one or more density modifiers (130), characterized in that the cells are grown in a culture media supplemented with from 2-20% human plasma. It also relates to a biological component (100) comprising: i. a matrix forming agent (120), ii. a plurality of cells (110), and iii. one or more density modifiers (130), wherein at performance competence, the cells are present in the matrix forming agent at a density of at least 3×107 cells/ml.

Abstract: The present invention describes new mammalian expression vectors comprising a novel combination of regulatory elements and one or more selection marker gene(s). The vector allows for incorporation of at least one, preferably two or more genes of interest, its/their subsequent expression, and for selection of transfected cells using, e.g., G418 and/or MTX. The pDGP?GOI vector as an example for a mammalian expression vector according to the present invention exhibits a 9555 bp sequence, one strand of which is represented by SEQ ID NO:2.

Abstract: The present invention relates to a method for increasing the expression of a protein in cells, preferably in eukaryotic cells, by reducing the number of RNase L cleavage sites in the coding and/or non-coding region of the nucleic acid sequence of said protein. Furthermore, it relates to nucleic acid sequences exhibiting a reduced number of RNase L cleavage sites as well as to the proteins translated from such sequences.

Abstract: Methods for enhancing iron uptake in cell culture are described. The methods include using a NTBI uptake activator and non-protein bound iron source. Suitable NTBI activators are delineated. Also described are serum-free culture media which can be used in the present methods.

Abstract: Disclosed herein are polypeptides, polynucleotides encoding, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.

Abstract: The use of a non-glycanated form of a polypeptide including an amino acid sequence having at least 90% amino acid identity with an amino acid sequence selected from the group consisting of SEQ ID No 1 and SEQ ID No 2 for manufacturing a medicament for preventing or treating a cancer.

Abstract: The present inventors developed hepatitis C virus 2b/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and the complete NS2 were replaced by the corresponding genes of the genotype 2b reference strain J8. Sequence analysis of recovered 2b/2a recombinants from 2 transfection experiments revealed that 2b/2a was genetically stable. Conclusion: The developed 2b/2a viruses provide a robust in vitro tool for research in HCV genotype 2b, including vaccine studies and functional analyses.

Abstract: Disclosed herein are methods for producing liver precursor cells as well as hepatocyte cells form pluripotent and/or multipotent cells. Also disclosed herein are methods of enriching isolating and/or purifying liver precursor cells and/or hepatocyte cells. Further disclosed are compositions comprising cell cultures and cell populations that are enriched for liver precursor cells or hepatocyte cells.

Abstract: Genotype 7a has been identified recently, thus not much is known about the biology of this new, major HCV genotype. The present inventors developed hepatitis C virus 7a/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and the complete NS2 were replaced by the corresponding genes of the genotype 7a strain QC69 and characterized them in Huh7.5 cells. Sequence analysis of 7a/JFH1 recombinants recovered after viral passage in Huh7.5 cells following 4 independent transfection experiments revealed adaptive mutations in Core, E2, NS2, NS5A and NS5B. In reverse genetic studies the importance of these mutations for improved growth kinetics was shown. Adapted 7a/JFH1 viruses showed growth kinetics, infectivity and RNA titers comparable to a previously developed 3a/JFH1 reference virus. Conclusion: The developed 7a/JFH1 viruses provide a robust in vitro tool for research in HCV genotype 7, including vaccine studies and functional analyses.

Abstract: An HCV/GBV-B chimeric virus which maintains the replication function of HCV and is capable of infecting tamarin is disclosed in order to construct an HCV animal model which can be used as a development or evaluation system for therapeutic agents for HCV. The HCV/GBV-B chimeric RNA comprises an RNA of hepatitis C virus and an RNA of GB virus-B, wherein the RNA of hepatitis C virus comprises an RNA encoding leucine at the 1804th position and lysine at the 1966th position in the amino acid sequence of the polyprotein of hepatitis C virus.

Abstract: Culture media comprising manganese and methods of culturing cells to improve sialylation and glycosylation of glycoproteins are provided.

Abstract: An objective of the present invention is to provide polynucleotides encoding insect desiccation resistance proteins, and uses thereof cDNA libraries were produced from Polypedilum vanderplanki larvae in a desiccated state, a P. vanderplanki EST database was constructed, and genes encoding LEA proteins were isolated. This resulted in the successful isolation of three types of novel gene encoding LEA-like proteins (PvLEA1, PvLEA2, and PvLEA3.) When secondary structure predictions and motif searches were performed on the proteins deduced from each of the genes, all three proteins had ?-helix-rich structures and LEA_4 motifs, which are characteristic of LEA proteins. Moreover, the recombinant proteins synthesized from PvLEA1, 2 and 3 genes were heat soluble even when boiling, so that PvLEA1, 2 and 3 proteins have highly hydrophilic property as well as plant LEA proteins. Therefore, the three isolated genes were found to be novel P. vanderplanki-derived LEA genes.

Abstract: A protein scaffold based on a consensus sequence of fibronectin type III (FN3) proteins, such as the tenth FN3 repeat from human fibronectin (human Tenascin), including isolated nucleic acids that encode a protein scaffold, vectors, host cells, and methods of making and using thereof, exhibit enhanced thermal and chemical stability while presenting six modifiable loop domains which can be engineered to form a binding partner capable of binding to a target for applications in diagnostic and/or therapeutic compositions, methods and devices.

Abstract: A protein inhibitor from Sulfolobulus Solfataricus and peptides thereof, which both have the ability to inhibit AARE and elastase. Similarly to the eukaryotic counterparts, the recombinant protein is able to inhibit in vitro the bovine alfa-chymotrypsin with a high specificity, and the porcine elastase but not all commercial available trypsins, features which distinguish all the members belonging to the family PEBP. Through site-specific mutagenesis techniques of the gene codifying SsCEI, it has been recognized the “reactive site loop”—RCL—on the inhibitor, responsible for the interaction with the eukarial protease target already identified. The inhibitor and the peptides thereof can be used as new compounds capable of modulating cognitive enhancement cardiovascular diseases, cancer, inflammation, hematological diseases, neurological diseases and urological diseases.

Abstract: A method of inhibiting graft rejection includes isolating Hepatic Stellate Cells from a mammal liver, activating the isolated Hepatic Stellate Cells, and administering a combination of Hepatic Stellate cells and a graft to a mammal.

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: In order to provide a method for producing neural stem cells easily and quickly by inducing differentiation of somatic cells directly into neurospheres, dedifferentiation factors are introduced into somatic cells, which are then cultured in suspension in the presence of growth factors to produce the neurospheres, thereby allowing the neural stem cells to be produced quickly without establishing iPS 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: The invention provides compositions and methods for detecting the presence of SARS-coronavirus, for screening anti-SARS coronavirus agents and vaccines, and for reducing infection with plus-strand RNA viruses such as SARS-coronavirus.

Abstract: The present invention relates to a human hepatoma-derived cell line, LH86, liver cell cultures, non-human animal models, artificial livers, liver assist devices, screening assays, and other applications of the cell line as an investigational tool.

Abstract: To provide a cell culture kit including cultured living cells of various donors, and a manufacturing method thereof. The cell culture kit includes a culture plate and living cells cultured thereon. The culture plate includes a plurality of microchambers (33) and living cells derived from various donors are adhered to surfaces of the plurality of microchambers (33). Specifically, living cells D1, D2, and D3 derived from various donors are adhered to the plurality of microchambers (33). In each microchamber (33), living cells derived from one donor or living cells derived from various donors may be cultured. The living cells derived from various donors are adhered and cultured in the cell culture kit as a whole, which makes it possible to provide a cell culture kit to conduct a test using cells derived from various donors.

Abstract: The present invention provides cell populations that are enriched for mesendoderm and mesoderm, and cell populations that are enriched for endoderm. The cell populations of the invention are useful for generating cells for cell replacement therapy. The present invention further provides a method of generating hepatocytes, cell populations enriched for hepatocytes, and a method of hepatocyte replacement therapy.

Abstract: Pathogenic effector proteins include one or more virulence motifs of amino acid consensus sequence BXZ, where B=RK or H; X=any amino acid or is absent; Z=L, M, I, W, Y or F) which bind to target polar lipids on a host (plant or animal) cell as a prerequisite for translocation of the pathogenic effector proteins into the cell. Translocation is prevented by binding blocking compounds to one or more motifs of the effector protein or to the lipid ligands of the host cell. The blocking compounds include synthetic or naturally occurring polypeptides which bind the polar lipids or the motifs, various polar lipids, the hydrophilic head-groups of polar lipids, etc. Suitable blocking compounds can be identified by assays demonstrating binding to the motifs or to the target polar lipids.

Abstract: To provide a cell storage method to store living cells while maintaining the functions of the living cells. An aspect of the cell storage method is a method to store living cells (40) by using a cell culture chamber (20) including a plurality of the microchambers (11), the method including: culturing the living cells by being adhered to the surfaces of a plurality of micro spaces; and after the culturing, pouring the culture medium (50) into the cell culture chamber (20) so as to cover the plurality of microchambers (11), and storing the living cells. The living cells are adhered to a cell culture chamber of an appropriate size and cultured to form a three-dimensional structure. This enables maintaining the three-dimensional structure of the living cells and storage of the living cells while maintaining the functions thereof.

Abstract: The present invention relates to a chimeric regulatory sequence with liver cell specificity. The chimeric regulatory sequence includes a proximal regulatory sequence and a distal enhancer 5? flanking region of human ?-fetoprotein (AFP) gene. The chimeric regulatory sequence is useful in purified specific lineages, such as liver cells, from other cell lineages.

Abstract: The present invention provides immortalized fetal liver cells that express the oncogene Simian virus (SV 40) large T-antigen and specific hepatocyte markers and a method for producing same.

Abstract: Methods and kits for expanding the number of hematopoietic stem cells are provided. The methods comprise incubating cells in medium comprising isolated IGFBP-2 and an angiopoietin-like protein (Angptl). Expanded HSCs are provided as well as culture media and kits for the expansion of human HSCs in a defined medium. Methods of administering expanded human HSCs to and individual are provided as well as methods of treating an individual by administering certain growth factors and cytokines.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of C-reactive protein. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding C-reactive protein. Methods of using these compounds for modulation of C-reactive protein expression and for treatment of diseases associated with expression of C-reactive protein are provided.

Abstract: The present invention is directed to methods of treating cells infected with HCV and mammals suffering from HCV infection by transfecting the infected cells with miRNA-196 mimic. miRNA-196 mimic significantly down-regulates Bach1 protein and HCV gene expression, while also up-regulating HMOX1 gene expression. miRNA-196 binds with the 3?-UTR of Bach1 mRNA to reduce the expression of Bach1. As such, miRNA-196 can play an important role in the regulation of HCV replication and HMOX1/Bach1 expression in hepatocytes. The present invention also provides a formulation for the treatment of cells expressing HCV comprising a therapeutically effective amount of miRNA-196 such that Bach1 and HCV gene expression are down-regulated while HMOX1 expression is increased. The formulations are adapted to enable the transfection of miRNA-196 mimic into hepatocytes expressing HCV proteins.

Abstract: A rotating wall vessel is used as a culture vessel and bioreactor for the cultivation of hepatocytes in the form of spheroids to generate a culture with many properties of the intact liver. These properties include enzyme activity comparable to fresh cells and long-term maintenance of viability and cellular function for periods on the order of months. The cultures may be used to produce hepatocyte products, evaluate metabolism of an agent, propagate Hepatitis C virus and test agents as inhibitors of this virus. Thus, the culture system disclosed herein makes long term functional cultivation of human hepatocytes feasible.

Abstract: The present invention provides a human hepatic stem cell which is sorted based on the presence or absence of the expression of at least one marker selected from the group consisting of CD318, CD90, CD66 and CD13. A method of preparing the human hepatic stem cell, a method of inducing differentiation of the same and a method of using the same are also provided.

Abstract: The present invention discloses novel methods and omental, myocardial, liver, lung, renal, peritoneal, intestinal and pancreatic mesothelial cells which are useful for a number of procedures including drug discovery, co-culturing, cell therapy and bioassay. The invention provides a method for isolating these cells that improves upon the methods previously used and provides cells isolated in quantity. The present invention provides a list of secreted proteins from omentum mesothelial cells that can be utilized in the described cell based assays.

Abstract: The present invention provides compositions comprising desiccated biologics comprising a cell, protein, virus, nucleic acid, carbohydrate, or lipid, or any combination thereof, along with at least one membrane penetrable sugar, and at least one membrane impenetrable sugar, wherein the moisture content is from 5% to 95%, and to methods of preparing the same, and to methods of treating animals using the same.

Abstract: The present invention relates to methods of identifying, collecting and isolating hematopoietic stem cells (HSCs) and compositions of purified HSCs. Specifically, the present invention provides methods of isolating and purifying CD150+ HSCs, CD48? HSCs, and CD244? HSCs. The present invention also relates to purified cell samples with enriched CD150+ HSCs, CD48? HSCs, and CD244? HSCs populations, as well as methods of treating subjects with such compositions.

Abstract: A method is provided for controlling the survival, proliferation, and/or differentiation of hepatic progenitors in vitro by using specific types of mesenchymal feeder cells or one of more of the paracrine signals produced by those feeders.

Abstract: Disclosed are methods for inducing cardiomyogenic differentiation in cells that are competent for differentiation along the cardiomyogenic lineage such as certain unfractionated bone marrow mononuclear cells (BMMNCs). In some embodiments, the methods include contacting a plurality of unfractionated, density gradient-separated BMMNCs with a cardiomyocyte differentiation-inducing amount of a Wnt11 gene product for a time and under conditions sufficient to induce cardiomyocyte differentiation in at least a subset of the BMMNCs. Also provided are methods for treating an injury to cardiac tissue in a subject using cells that have been induced to differentiate along the cardiomyogenic lineage, recombinant host cells comprising an expression vector that encodes a Wnt11 polypeptide or a functional fragment thereof, and systems for inducing cardiomyogenic differentiation in a cultured cell.

Abstract: Provided is a cell culture method whereby an in vivo function can be sustained over a long period of time and culture can be conducted using the minimum number of cells required. The cell culture method includes culturing undifferentiated cells in a layered state in a partitioned micro-space and obtains differentiated cells. When screening a pharmaceutical agent, undifferentiated cells capable of differentiating into liver cells, intestinal epithelial cells, nerve cells, myocardial cells and vascular endothelial cells are preferred. Particularly, in the prediction of pharmacokinetics or the like for humans, human cells are preferred.

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: The invention relates to peptide compositions for use in the preparation of pharmaceutical compositions against hepatitis C virus, as well as corresponding methods.

Abstract: Human adult hepatocytes are transplanted into an immunodeficient hepatopathy mouse and then human growth hormone is administered to the mouse to thereby elevate twice or more the replacement ratio by the human adult hepatocytes having been transplanted. Further, human growth hormone is administered to an immunodeficient hepatopathy mouse carrying human young hepatocytes transplanted thereinto so as to improve fatty liver of the mouse in which about 70% or more of the hepatocytes have been replaced by the human hepatocytes.

Abstract: Methods of transplanting cells, such as hepatocytes, are presented herein. Such methods are useful for treating liver disease as well as other disorders.

Abstract: Described herein is a method of expanding human hepatocytes in vivo using an immunodeficient mouse which is further deficient in fumarylacetoacetate hydrolase (Fah). The method comprises transplanting human hepatocytes into the immunodeficient and Fah-deficient mice, allowing the hepatocytes to expand and collecting the expanded human hepatocytes. The method also allows serial transplantation of the human hepatocytes into secondary, tertiary, quaternary or additional mice. Also provided are mutant mice comprising homozygous deletions or point mutations in the Fah, Rag2 and Il2rg genes.

Abstract: The present invention relates, in one aspect, to a vector comprising (a) a first polynucleotide capable of encoding a first (poly)peptide comprising at least one cysteine residue, and (b) a second polynucleotide capable of encoding a second (poly)peptide which is a cell surface anchor comprising at least one cysteine residue, wherein the vector is operable in a eukaryotic host cell to express and to cause or allow the attachment of said first (poly)peptide to said second (poly)peptide by formation of a disulfide bond between said cysteine residues comprised within said first (polypeptide and said second (poly)peptide, respectively, wherein said first (poly)peptide is exhibited at the surface of a eukaryotic host cell.

Abstract: Disclosed herein are methods and compositions for inactivating a glutamine synthetase (GS) gene, using fusion proteins comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding said fusion proteins are also provided, as are cells comprising said polynucleotides and fusion proteins.

Abstract: Clostridial toxin substrates comprising a lanthanide donor complex, an acceptor, and a Clostridial toxin recognition sequence including a cleavage site; methods for determining the activity of a Clostridial toxin from a test sample using such Clostridial toxin substrates; cell compositions comprising such Clostridial toxin substrates and a Clostridial toxin receptor; and methods for determining the activity of a Clostridial toxin from a test sample using such cell compositions.

Abstract: The present invention relates to a composition for preserving cells, tissues and organs, comprising as an active ingredient indole and indazole compounds of formula (1), or a pharmaceutically acceptable salt or isomer thereof, which are effective for preventing injury of organs, isolated cell systems or tissues caused by cold storage, transplant operation or post-transplantation reperfusion; a preservation method; and a preparation method of the composition.

Abstract: This invention relates to a nucleic acid comprising a nucleic acid sequence encoding an epitope tag peptide in the hypervariable region 1 of the E2 protein of naturally occurring or chimeric hepatitis C virus (HCV) genome, an epitope-tagged HCV particle encoded by such nucleic acid, and a method for purifying the HCV particles at a high purity in a simple manner with the use of an anti-epitope tag antibody-immobilized support.

Abstract: The present invention relates to precursor cells to hepatic stellate cells, compositions comprising same and methods of isolating same. The surface antigenic profile of the precursors is MHC class Ia negative, ICAM-1+, VCAM-1+, ?3-integrin+. In addition to expression of these surface markers, the cells also express the intracellular markers desmin, vimentin, smooth muscle ?-actin, nestin, hepatocyte growth factor, stromal derived factor-1? and H1x homeobox transcriptional factor.