Abstract: Provided herein are synthetic, three-dimensional (3D) bioprinted tissue constructs comprising porcine cells and methods of producing and using the same. The synthetic 3D bioprinted tissue constructs are fabricated by bioprinting spheroids comprising porcine cells, including genetically engineered cells, on a microneedle mold and fusing the spheroids to form an engineered tissue construct. Also provided are methods of using scaffold-free 3D bioprinted tissue constructs for applications related to drug screening and toxicity screening.

Abstract: Methods are provided for producing a population of hepatocyte-like cells (iHeps) from a population of adipocyte-derived stem cells (ASCs). Aspects of the methods include placing a population of ASCs into a three dimensional culture (e.g., hanging drop suspension culture, high density culture, spinner flask culture, microcarrier culture, etc.), and contacting the cells with a first and second culture medium. Also provided are methods of treating an individual, which include producing a population of iHeps from a population of ASCs, and administering an effective number of iHeps into the individual. Kits for practicing the methods are also described herein.

Abstract: The invention relates to a liver organoid, uses thereof and method for obtaining them.

Abstract: The present invention relates to directed differentiation and maturation of mammalian hepatocytes, such as human hepatocytes. The hepatocyte obtained in accordance with the present invention show a phenotype which is more similar to that of primary hepatocytes than previously shown. In particular, the present invention relates to exposure of mammalian hepatocytes, such as human hepatocytes, to at least one maturation factor selected from the group consisting of Src kinase inhibitors, vitamin D including precursors, metabolites and analogs thereof, hypoxia inducing compounds, sphingosine and sphingosine derivatives, activators of peroxisome proliferator-activated receptors (PPARs), platelet-activating factor (PAF), PKC inhibitors, and combinations thereof.

Abstract: The present disclosure provides methods, and related kits, for directing cell attachment and spreading on a substrate and inducing isotropic spreading of cells; provides methods, and related kits, for cell sorting; and further provides methods, and related kits, for guided induction of stem cell differentiation.

Abstract: The present disclosure relates to the design of a retro- or lenti-viral HBV (hepatitis B virus) hybrid vector for delivery of HBV genomes into cells such as liver cells. Provided herein are design, methodology, compositions, and the like for such vectors for delivering HBV genes or genomes into cells for anti-HBV drug screening, HBV research, or test of anti-HBV therapeutics.

Abstract: It is an object of the present invention to provide a method for producing frozen mesenchymal cells maintaining a viability and an adhesion rate similar to those of cells obtained through a step of culture, without undergoing a step of culturing cells before cryopreservation step. The method comprises dissociating cells by treating a tissue with proteolytic enzymes and cryopreserving the cells. The present invention provides a method for producing frozen mesenchymal cells maintaining a viability and an adhesion rate similar to those of culturing cells without a step of culturings cells before cryopreserving.

Abstract: Provided herein are compositions, methods and uses that relate to or result from the identification of markers that can distinguish between cells at different stages of pluripotency. Certain embodiments provide markers that can distinguish between parental cells (i.e. differentiated cells), partially pluripotent (i.e. partially reprogrammed) and pluripotent (i.e. fully reprogrammed cells). Also provided here are uses of such differential markers, for example, in identification of cell potential, in diagnostics, in differential separation, and in creating efficient workflows that involve fewer steps and lesser time in identifying or separating a desired reprogrammed clone or cell line from a mixture of cells at various stages of pluripotency. In certain embodiments, the activity of these markers can be manipulated to influence cell potential for research or medical purposes.

Abstract: This invention relates to the in vitro differentiation of pluripotent cells into pancreatic progenitors by i) culturing pluripotent cells in a definitive endoderm (DE) medium comprising a TGFp ligand, fibroblast growth factor (FGF), bone morphogenetic protein (BMP), a PI3K inhibitor and optionally a GSK3 ? inhibitor to produce a population of definitive endoderm cells, ii) culturing the definitive endoderm cells in a first pancreatic medium comprising an activin antagonist; FGF; retinoic acid; and a BMP inhibitor to produce a population of dorsal foregut cells; iii) culturing the dorsal foregut cells in a second pancreatic medium comprising FGF, retinoic acid, a BMP inhibitor, and a hedgehog signalling inhibitor, and; iv) culturing the endoderm cells in a third pancreatic medium comprising FGF. The progenitor cells thus produced may be further differentiated into pancreatic endocrine cells. These methods may be useful, for example, in producing pancreatic cells for therapy or disease modelling.

Abstract: The present invention discloses cryopreserved recombinant cells for screening drug candidates that transiently overexpress one or more drug transporter proteins and/or drug metabolizing enzymes. Advantageously, such cells provide a cost-efficient consumable product that streamlines the process of screening whether drug candidates are substrates or inhibitors of drug transporter proteins and/or drug metabolizing enzymes.

Abstract: The invention relates to a liver organoid, uses thereof and method for obtaining them.

Abstract: A method of repairing diseased or dysfunctional pancreas or liver is provided. The method involves preparation of a suspension of stem cells and/or progenitor cells such as biliary tree stem cells, hepatic stem cells, pancreatic stem cells or their descendants, committed progenitor cells, from healthy tissue of the patient or of the biliary tree of a non-autologous donor and engrafting the cells into the wall of bile ducts near to the organ to be treated. The graft consists of stem cells or progenitors that are admixed with biomaterials and, optionally, with cytokines and/or native epithelial-mesenchymal cells appropriate for the maturational lineage stage of the cells to be engrafted.

Abstract: The invention relates to a new type of mesenchymal stem cells (MSC) which co-express at least one mesenchymal marker, preferably at least CD105 and CD34. Also provided are bone-forming cells having an analogous phenotype. The invention also provides the cells and cell populations, as well as further products comprising such and uses thereof in bone therapy.

Abstract: The present invention relates to a method for diagnosing a carcinoma or a residual disease associated thereto, or for the prognosis of a carcinoma, or for monitoring the effectiveness of an anti-tumour therapy directed against a carcinoma, or for monitoring the follow-up of an individual affected by a carcinoma, in particular colorectal carcinoma, carcinoma of the stomach, mammary carcinoma, pulmonary carcinoma or carcinoma of the prostate, carcinoma of the liver, carcinoma of the ovary, carcinoma of the kidney, carcinoma of the thyroid, carcinoma of the bladder or carcinoma of the pancreas. The method of the invention consists in placing adult stem cells in contact with a sample of a haemo-derivative of the individual to be analyzed and in verifying the expression of at least an epithelial marker in the stem cells by means of immunofluorescence, immunohistochemistry, ELISA or RT-PCR.

Abstract: Conventional CTC detection methods have been problematic in that 1) there is no technique for automatically determining and counting live CTCs in a brief period of time, 2) no process has been developed for detecting, counting, and thereafter collecting and culturing live CTCs, and 3) there exists no flow cytometer that is contamination free and is capable of measuring an entire sample. Provided is a CTC detection method which comprises a pre-treatment step for concentrating and fluorescence staining CTCs, and a step for identifying and counting CTCs. The pre-treatment step includes attaching magnetic beads to EpCAM antibodies expressed by epithelial cell-derived CTCs and concentrating the CTCs through the use of a magnet, fluorescently labeling an epithelia cell surface marker of the CTCs through the use of EpCAM antibodies or 5E11 antibodies, and performing two types of nuclear staining, one being cell membrane-permeable and the other being cell membrane-impermeable.

Abstract: Methods, compositions and kits for producing functional neurons, astroctyes, oligodendrocytes and progenitor cells thereof are provided. These methods, compositions and kits find use in producing neurons, astrocytes, oligodendrocytes, and progenitor cells thereof for transplantation, for experimental evaluation, as a source of lineage- and cell-specific products, and the like, for example for use in treating human disorders of the CNS. Also provided are methods, compositions and kits for screening candidate agents for activity in converting cells into neuronal cells, astrocytes, oligodendrocytes, and progenitor cells thereof.

Abstract: Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles.

Abstract: Methods for de-differentiating or altering the life-span of desired “recipient” cells, e.g., human somatic cells, by the introduction of cytoplasm from a more primitive, less differentiated cell type, e.g., oocyte or blastomere are provided. These methods can be used to produce embryonic stem cells and to increase the efficiency of gene therapy by allowing for desired cells to be subjected to multiple genetic modifications without becoming senescent. Such cytoplasm may be fractionated and/or subjected to subtractive hybridization and the active materials (sufficient for de-differentiation) identified and produced by recombinant methods.

Abstract: The present invention relates to an repebody capable of binding specifically to interleukin-6 (IL-6) to inhibit the biological activity of IL-6, a polynucleotide encoding the repebody, a vector comprising the polynucleotide, a recombinant microorganism having introduced therein the polynucleotide or the vector, a method of producing the repebody using the recombinant microorganism, a composition for preventing or treating cancer, which comprises the repebody, and a method for preventing or treating cancer, which comprises administering the composition for preventing or treating cancer, which comprises the repebody. The repebody of the present invention significantly reduces the activity of STAT3 and the concentration of interleukin-6, and thus can be widely used as an agent for preventing or treating IL-6-related diseases.

Abstract: Replicons of genotype 4d hepatitis C virus (HCV) are provided. These replicons contain adaptive mutations giving rise to the HCV's capability to replicate in vitro. Methods of preparing genotype 4d replicons and methods of using these replicons to screen antiviral agents are also provided.

Abstract: Human hepatocyte cell cultures and their use in bioreactors and bioartificial liver (BAL) systems are provided. The cells have constitutive liver-specific metabolic activity resembling that of freshly isolated human hepatocytes. The hepatocyte cells and BAL systems can be used to treat patients suffering from acute liver failure, end-stage liver disease, or acute-on-chronic liver disease.

Abstract: Analyte sensors, methods for producing and using analyte sensors, methods of detecting and/or measuring analyte activity, detecting pH change, and/or, controlling the concentration of an analyte in a system, are disclosed. Embodiments of the analyte sensors according to the disclosure can provide an accurate and convenient method for characterizing analyte activity, detecting pH change, controlling the concentration of an analyte in a system, and the like, in both in vivo and in vitro environments, in particular in living cell imaging.

Abstract: The invention provides improved adeno-associated virus (AAV) Factor VIII (FVIII) vectors, including AAV FVIII vectors that produce a functional Factor VIII polypeptide and AAV FVIII vectors with high expression activity.

Abstract: Disclosed herein are compositions for linking DNA binding domains and cleavage domains (or cleavage half-domains) to form non-naturally occurring nucleases. Also described are methods of making and using compositions comprising these linkers.

Abstract: The present invention relates to a one-vector expression system comprising a sequence encoding two polypeptides, such as tyrosine hydroxylase (TH) and GTP-cyclohydrolase 1 (GCH1). The two polypeptides can be should preferentially be expressed at a ratio between 3:1 and 15:1, such as between 3:1 and 7:1. The invention is useful in the treatment of catecholamine deficient disorders, such as dopamine deficient disorders including but not limited to Parkinson's Disease. Moreover, the present invention provides a method to deliver the vector construct in order to limit the increased production of the catecholamine to the cells in need thereof.

Abstract: The method of the invention provides for producing a heterologous protein in mammalian host cells having nucleic acid encoding Hepatitis B X protein and the heterologous protein, by growing mammalian host cells selected from the group consistng of HKB11, CHO, BHK21, C2C12, and HEK293 cells, by growing mammalian host cells in non-adherent suspension culture, or by growing mammalian host cells which contain nucleic acid providing exogenous X-box Binding Protein, XBP1s. The conditions should be such that HBx, exogenous XBP1s if present, and the heterologous protein are expressed by the mammalian cells. The invention includes compositions for carrying out the method.

Abstract: The present invention relates generally to a population of cells genetically modified to produce insulin in a glucose responsive manner and uses thereof. More particularly, the present invention relates to a population of cells genetically modified to produce insulin in response to physiologically relevant levels of glucose and uses thereof. The cells of the present invention are useful in a wide variety of applications, in particular in the context of therapeutic and prophylactic regimes directed to the treatment of diabetes and/or the amelioration of symptoms associated with diabetes, based on the transplantation of the cells of the present invention into mammals requiring treatment. Also facilitated is the design of in vitro based screening systems for testing the therapeutic effectiveness and/or toxicity of potential adjunctive treatment regimes.

Abstract: Disclosed are methods of gene delivery using capsid-modified recombinant adeno-associated viral (rAAV) vectors. Exemplary methods are provided employing vectors that have altered affinity for heparin or heparin sulfate, as well as vectors, expression systems, and rAAV virions that lack functional VP2 protein expression, but are nevertheless, fully virulent. Also provided by the invention are methods employing the rAAV vector-based compositions, virus particles, host cells, and pharmaceutical formulations in the expression of selected therapeutic proteins, polypeptides, peptides, antisense oligonucleotides and/or ribozymes in selected mammals, including organs, tissues, and human host cells.

Abstract: The invention describes novel virus-like particles for use as vaccines, diagnostic tools and R&D tools based on recombinant DNA and cell cultivation techniques for production. The recombinant virus-like particles of the invention are assembled by polypeptide chains that incorporate several, in particular two or more, different epitopes which are selected either (a) from different viral strains of the same virus and/or (b) from different serotypes of the same virus and/or (c) from different viral strains specific for different hosts. These epitopes are then displayed on the particle surface.

Abstract: The present invention relates to the preparation of poly(amino oxalate) (PAOX) using oxalyl chloride, 1,4-cyclohexanedimethanol, and piperazinediethanol, the preparation of biodegradable polymer particles using the PAOX, and the use of PAOX particles as a drug delivery vehicle. The PAOX according to the present invention is a polymer that has three characteristics of biodegradability, biocompatibility, and cationic properties at the same time with appropriate hydrophobicity and thus can be prepared as particles that allow rapid drug release. Moreover, the particles improve the delivery efficiency of a drug into cells and thus can be efficiently used as a drug delivery vehicle for the treatment of acute inflammatory diseases such as acute liver failure and acute lung injury.

Abstract: Provided is a cell cultivation method in which the cell is cultured using a peptide hydrogel as a scaffold, for carrying out high-dimensional culture of a cell such as porcine hepatocyte, human hepatocyte, porcine pancreatic islet or human pancreatic islet for a long period under conditions where cell survival, cell morphology and cell functions are maintained. Also provided are a cell culture including a cell and a peptide hydrogel obtained by the above-described cultivation method, a bioreactor including the cell culture, and a cell preparation including the cell culture.

Abstract: The present invention relates to a method for preparing spheroids of human primary hepatocytes. The method comprises culturing of isolated human primary hepatocytes on a polysaccharide scaffold under conditions that allow the formation of hepatocyte spheroids, and subsequently dissolving the polysaccharide scaffold to release the hepatocyte spheroids. The spheroids obtained by the method of the invention are particularly suitable for being transplanted into a subject afflicted with a liver disease.

Abstract: A method for measuring human CYP3A inducibility upon administration of a test drug, characterized in that a non-human animal to which a test drug is administered or a population of human cells cultured in a medium containing a test drug is infected with viruses (A) and (B); virus (A) being an adenovirus which is used as a vector and engineered by incorporating thereto a detectable reporter gene and at least 3 human PXR binding regions falling within an untranslated region of a human CYP3A gene, and virus (B) being an adenovirus which is used as a vector and engineered by incorporating thereto a human PXR cDNA; and subsequently expression level of the reporter gene is determined in the non-human animal or the cultured human cells.

Abstract: This document provides methods and materials related to differentiating iPS cells into glucose-responsive, insulin-secreting progeny. For example, methods and material for using indolactam V (ILV) and glucagon like peptide-1 (GLP-1) to produce glucose-responsive, insulin-secreting progeny from iPS cells are provided.

Abstract: The present invention relates to a nucleic acid containing at least one homing endonuclease site (HE) and at least one restriction enzyme site (X) wherein the HE and X sites are selected such that HE and X result in compatible cohesive ends when cut by the homing endonuclease and restriction enzyme, respectively, and the ligation product of HE and X cohesive ends can neither be cleaved by the homing endonuclease nor by the restriction enzyme. Further subject-matter of the present invention relates to a vector comprising the nucleic acid of the present invention, host cells containing the nucleic acid and/or the vector; a kit for cloning and/or expression of multiprotein complexes making use of the vector and the host cells, a method for producing a vector containing multiple expression cassettes, and a method for producing multiprotein complexes.

Abstract: Replicons of genotype 4 hepatitis C virus (HCV) are provided. These replicons contains adaptive mutations giving rise to the HCV's capability to replicate in vitro. Methods of preparing genotype 4 replicons and methods of using these replicons to screen antiviral agents are also provided.

Abstract: Replicons of genotype 3 of hepatitis C virus (HCV) are provided. These replicons contains adaptive mutations giving rise to the HCV's capability to replicate in vitro. Methods of preparing genotype 3 replicons and methods of using these replicons to screen antiviral agents are also provided.

Abstract: The invention relates to an expression cassette including a gene of interest under the control of an inducible promoter, characterized in that said inducible promoter includes at least one CARE regulatory sequence (C/EBP-ATF responsive element) and a minimal promoter. The invention also relates to a vector and a host cell, as well as to a pharmaceutical composition including such a cassette, and to the use thereof for treating diseases by gene therapy.

Abstract: The invention relates to the discovery that DHH derived from stem cells are permissive for infection by hepatitis viruses (HV), such as hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV) and hepatitis E virus (HEV). Included in the invention are HV-permissive DHHs and methods of making an HCV-permissive DHH derived from a stem cell. Also included is an HV culture system comprising at least one HV-permissive DHH. The HV-permissive DHH and HV culture system are useful for conducting HV life cycle analyses, diagnosing a subject as being infected with HV, genotyping and characterizing the HV of a subject infected with HV, detecting drug resistance of HV obtained from a subject infected with HV, screening for and identify modulators of HV infection, and monitoring the effect of a treatment of HV in a subject.

Abstract: The present invention provides RNAi agents, e.g., double stranded RNAi agents, that target the transthyretin (TTR) gene and methods of using such RNAi agents for treating or preventing TTR-associated diseases.

Abstract: The present provides fusion proteins comprising PDGF and VEGF binding portions, and recombinant viral particles encoding the fusion proteins. Compositions comprising the fusion proteins and viral particles as well as methods of using the same are also provided.

Abstract: Synthetic cell culture surfaces, including a hydrophobe modified cellulose or an hydroxylated acrylate polymer composition and optionally including a silica source, cell culture coating and cell culture articles incorporating the composition, and methods of making and using the articles for cell culture, as defined herein.

Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.

Abstract: The present invention relates to a method of producing cells having at least one characteristic of human hepatocytes as well as to cells produced by said method and uses of these cells.

Abstract: A subgenomic replicon RNA (nucleic acid) having an excellent autonomously replicating ability and a fullgenomic replicon RNA (nucleic acid) having an excellent autonomously replicating ability and infectious HCV particle-producing ability, each derived from a novel HCV of genotype 1b, are provided. Particularly, a subgenomic replicon RNA (nucleic acid) and a fullgenomic replicon RNA (nucleic acid), each derived from an HCV genome of the NC1 strain which is a novel HCV genotype 1b isolated from a patient with acute severe hepatitis C, are provided.

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: A nucleic acid includes the 5? untranslated region, a virus protein-coding region including the NS3 protein coding sequence, the NS4A protein coding sequence, the NS4B protein coding sequence, the NS5A protein coding sequence, and the NS5B protein coding sequence, and the 3? untranslated region of the HCV J6CF genome in that order from the 5? to 3? direction. The NS4A protein coding sequence has a mutation for substituting alanine at position 1680 with glutamic acid, as determined on the basis of the amino acid sequence of the precursor protein of the J6CF strain.

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

Abstract: The present invention relates to fluorescent proteins, in particular green fluorescent proteins (GFPs), with increased activity in cells, and thus increased signal strength. A further aspect of the present invention relates to the use of peptides for increasing the expression and/or stability of a protein in a cell.

Abstract: A bioartificial liver system is described that incorporates a cell reservoir and hepatocyte spheroids to both increase the number of and longevity of cells in the system. Additional methods are also described for forming spheroid aggregates from isolated hepatocytes.