Abstract: The present invention relates to a novel hepatocyte-like cell progenitor and/or a novel hepatocyte-like cell derived via definitive endoderm from human blastocyst-derived stem (hBS) cells, to a method for the preparation of such cells and to the potential use of such cells in e.g. pharmaceutical drug discovery and development, toxicity testing, cell therapy and medical treatment. In particular is presented a definitive endoderm derived hepatocyte-like cell with important liver-expressed marker genes and important metabolizing enzymes, as well as drug transporters.

Abstract: The present invention relates to the use of 3-dimensional (3D) synthetic or animal-derived bioscaffolds as substrates for the improved growth and differentiation of hPS (Human pluripotent stem cells); these scaffolds being adapted for use in conjunction with existing cell culture lab plastic-ware. More specifically, it relates to the seeding of these scaffolds, either alone or in conjunction with various biologic matrix coatings, with hPS cells for the improved differentiation of said hPS cells into hepatocyte or hepatocyte-like cell types. The invention also relates to the seeding of partially-differentiated hepatocyte progenitors onto scaffolds for further differentiation into more mature hepatocyte-cell types.

Abstract: The present invention relates to the use of 3-dimensional (3D) synthetic or animal-derived bioscaffolds as substrates for the improved growth and differentiation of hPS (Human pluripotent stem cells); these scaffolds being adapted for use in conjunction with existing cell culture lab plastic-ware. More specifically, it relates to the seeding of these scaffolds, either alone or in conjunction with various biologic matrix coatings, with hPS cells for the improved differentiation of said hPS cells into hepatocyte or hepatocyte-like cell types. The invention also relates to the seeding of partially-differentiated hepatocyte progenitors onto scaffolds for further differentiation into more mature hepatocyte-cell types.

Abstract: Provided are improved methods using Glycogen synthase kinase 3 (GSK3) inhibitors by which endodermal cells, notably endodermal cells derived from human pluripotent stem cells (hPS), such as but not limited to hiPS-cells and hES-cells may be differentiated into hepatocyte like cells. The specific modulation of wingless integration gene (WNT)-signalling pathway and use of GSK3 inhibitors achieve direct differentiation and maturation of hepatocytes derived from human pluripotent stem (hPS) cells. GSK-3 inhibitors, when added to the growth medium at certain developmental stages, leads to more mature and functional features for the hepatocyte like cells as well as more pure and homogenous populations of hepatocyte like cells. Provided are also hepatocyte like cells obtained by these methods as well as compositions comprising them.

Abstract: The present invention relates to a method to control differentiation of human pluripotent stem cells, including human balstocyst derived stem (hBS) cells and to obtain specific endoderm cells. In particular, present invention relates to the use of FGF2 as the key factor in a specific concentration to control differentiation of definitive endoderm cells derived from hPS cells to specific endoderm cells. The invention also provides methods of obtaining endoderm cells comprising the use of FGFR and activation of the MAPK signalling pathway.

Abstract: The present invention relates to the use of 3D culturing systems for the derivation of hepatocyte-like cells from human pluripotent stem cells (hPS). In particular, the invention concerns the directed differentiation and maturation of human pluripotent stem cells into hepatocyte like cells in 3D hollow fiber capillary bioreactors.

Abstract: An in vitro toxicity assay based on human blastocyst-derived stem cells for the detection of toxicity in the human species is provided, which enables novel detection of in vitro human toxicity for a substance and/or more efficiently detects human toxicity compared to non-human assays. Furthermore, the detection of toxicity for substances is enabled, which is known to display inter-species differences and the toxic effect was not detectable by toxicological tests in mice.

Abstract: The present invention discloses a novel antibody HES5:3:3 directed towards a specific antigen present on human pluripotent stem (hPS) cells and cancer tissue. The antibody can be used as a tool for human pluripotent stem (hPS) cell applications, such as the separation, surface adhesion and enhanced survival of said hPS cells. Furthermore, the present invention refers to the use of the antigen detection for cancer applications.

Abstract: The present invention relates to a novel hepatocyte-like cell population derived from hBS cells and to the potential use of such heopatocyte-like cells in e.g. medical treatment, drug screening and toxicity testing. Furthermore, the invention relates to 5 hepatoblast-like cells that may have suitable characteristics so that they can be used for the same applications as the hepatocyte-like cells and that furthermore may be used in in vitro studies of hepatogenesis such as early hepatogenesis or hepatoregenerative disorders. Both the hepatocyte-like and the hepatoblast-like cells according to the invention express drug transporter and/or drug metabolising 10 characteristics either at the gene or protein expression level.

Abstract: A cluster is provided comprising cardiomyocyte-like cells, wherein the cluster has i) contracting cells, ii) cells that are electrically connected, and expresses iii) cardiac markers including Nkx.2.5, troponin and myosin, iv) markers for functional adrenergic receptors, v) markers for functional muscarinic receptors, vi) markers for functional ion-channels including hERG, Na+, Ca2+ and K+ channels, vii) one or more endodermal markers selected from the group consisting of AFP, TF, APOA2, AHSG, SERPINA1, APOA1, APOC3, TTR1 APOB, and RBP4. A method for preparing the clusters and methods utilizing the clusters in drug discovery and toxicity screenings are described.

Abstract: The present disclosure relates to a novel hepatocyte-like cell progenitor and/or a novel hepatocyte-like cell derived via definitive endoderm from human blastocyst-derived stem (hBS) cells, to a method for the preparation of such cells and to the potential use of such cells in, e.g., pharmaceutical drug discovery and development, toxicity testing, cell therapy and medical treatment. In particular is presented a definitive endoderm derived hepatocyte-like cell with important liver-expressed marker genes and important metabolizing enzymes, as well as drug transporters.

Abstract: Technology is provided for the transfer of human blastocyst-derived stem cells (hBS cells) to a feeder-free culture system and propagation of the cells in such a feeder-free culture system, the method comprising the following steps of (a) transferring the blastocyst-derived stem cells from feeder to feeder free culture by mechanical treatment, (b) culturing the blastocyst-derived stem cells under feeder cell free growth conditions in a suitable growth medium and/or on a suitable support substrate, and (c) optionally passaging the blastocyst derived stem cell line every 3-10 days by enzymatic and/or mechanical treatment. The application of hBS cells cultured under a feeder-free condition in medicine (e.g., myocardial regeneration) and screening and toxicity testing also is provided.

Abstract: A combined scalable in vitro differentiation and assay system based on human blastocyst-derived stem (hBS) cells or cells derived from hBS cells is provided. This system makes it possible to merge both the differentiation and the assay parts of the system into one. The advantage of the combined assay system is that the hBS cells or the cells derived from hBS cells in the differentiation system are directly applicable for assays, in a large variety of assay units, such as different multiwell plates. The system offers a major improvement compared to the prior art, since the cells are differentiated in the same format as when further being subject to analysis. The starting cell material can be homogenously distributed across a variety of different plates, for use and can be cultured when attached, when semi-attached or when in suspension.

Abstract: A method for obtaining a stable xeno-free hBS cell line, xeno-free hBS cell lines obtained according to said method and use thereof.

Abstract: An in vitro toxicity assay based on human blastocyst-derived stem cells for the detection of toxicity in the human species is provided, which enables novel detection of in vitro human toxicity for a substance and/or more efficiently detects human toxicity compared to non-human assays. Furthermore, the detection of toxicity for substances is enabled, which is known to display inter-species differences and the toxic effect was not detectable by toxicological tests in mice.

Abstract: A novel population of multipotent cardiac precursor (MCP) cells derived from human blastocysts derived stem cells is disclosed, methods for the preparation thereof and use of the cells for in vitro testing. Basement cells derived from hBS cells are also disclosed and method for the preparation of MCP cells from basement cells. The MCP cells have the following characteristics i) at least 1% of the cells exhibit no antigen expression of one or more markers for undifferentiated cell, the marker being selected from the group consisting of SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4, ii) at least 1% of the cells exhibit no protein expression of one or more of a neural marker including nestin or GFAP iii) at least 1% of the cells exhibit protein and/or gene expression of one or more of a mesodermal marker including brachyury, vimentin or desmin iv) at least 1% of the cells exhibit protein and/or gene expression of Flk-1 (KDR). Furthermore, the MCP cells have a characteristic morphology.

Abstract: A method for the transfer of human blastocyst-derived stem cells (hBS cells) to feeder-free culture system and propagation of the cells in such a feeder-free culture system, the method comprising the following steps of (a) transferring the balstocyst-derived stem cells from feeder to feeder free culture by mechanical treatment, (b) optionally, culturing the blastocyst-derived stem cells under feeder cell free growth conditions in a suitable growth medium and/or on a suitable support substrate, and (c) optionally passaging the blastocyst derived stem cell line every 3-10 days by enzymatic and/or mechanical treatment. The invention also relates to the application of hBS cells cultured under feeder free condition in medicine (e.g., myocardial regeneration) and screening and toxicity tests.

Abstract: The present invention relates to a culture system for and a method for propagation of human blastocyst-derived stem cells (hBS cells) upon enzymatic dissociation into a single cell suspension. The culture system for propagation of human blastocyst-derived stem (hBS) cells comprises i) human feeder cells at a density of at least 50,000 cells/cm2, ii) one or more dissociation agents for dissociation of hBS cell colonies into a single cell suspension, and iii) a supportive culture medium, which culture system makes it possible to propagate hBS cells by dissociation of hBS cell colonies into a single cell suspension at each consecutive passage for an extended time period, while maintaining the significant characteristics of hBS cells.

Abstract: The present invention relates to a novel hepatocyte-like cell progenitor and/or a novel hepatocyte-like cell derived via definitive endoderm from human blastocyst-derived stem (hBS) cells, to a method for the preparation of such cells and to the potential use of such cells in e.g. pharmaceutical drug discovery and development, toxicity testing, cell therapy and medical treatment. In particular is presented a definitive endoderm derived hepatocyte-like cell with important liver-expressed marker genes and important metabolizing enzymes, as well as drug transporters.

Abstract: An improved method for vitrification of biological cells, especially blastocyst-derived stem cells (BS cells). The method is very mild for the cells that remain viable after they have been thawed. The method comprises, i) transfer of the cells to a first solution (solution A), ii) optionally incubation of the cells in the first solution, iii) transfer the cells obtained in step i) or ii) to a second solution (solution B), iv) optionally incubation of the cells in the second solution, v) transfer of the cells obtained from step iii) or iv) into one or more closed straws with dimensions that allow a volume of at least 20 ?l to be contained in them vi) sealing the one or more closed straws, and vii) vitrification of the one or more closed straws. An important feature of the present invention is the use of closed straw and that relatively large volumes can be efficiently vitrified and subsequently thawed.