Miltenyi Biotec B.v. Co., Kg

Kristin Becker

Andreas Bosio

Dominik Eckardt

Olaf Hardt

David Joel Agorku

Olaf Thorsten Hardt

The present invention provides a cell culture for obtaining an epithelial organoid, the cell culture comprising i) epithelial stem cells, or tissue fragments comprising said epithelial stem cells, ii) a basal medium for animal or human cells, iii) a Bone Morphogenetic Protein (BMP) inhibitor, iv) a mitogenic growth factor selected from the group consisting of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and keratinocyte growth factor (KGF), v) at least one soluble culture enhancer, wherein said at least one culture enhancer induces correct polarization of the cells in said cell culture within the developing organoid such as a laminin/entactin complex or entactin, and vi) a Wnt agonist if said epithelial stem cells, or tissue fragments comprising said epithelial stem cells are healthy cells, wherein said at least one soluble culture enhancer in said cell culture is a laminin/entactin complex in a concentration between 0.2 mg/ml and 3.4 mg/ml. A cell culture medium, an in-vitro method for obtaining an epithelial organoid, and an epithelial organoid obtained by said method are also disclosed.

Cell culture medium and method for generation of epithelial organoids from epithelial stem cells

The present invention provides a method for generation, isolation, detection and/or analysis of cardiomyocytes derived from a starting cell composition comprising pluripotent and/or multipotent stem cells, the method comprising the steps a) differentiating said pluripotent and/or multipotent stem cells into cardiovascular cells, thereby generating a sample comprising cardiomyocytes and non-cardiomyocytes; and b) labeling the non-cardiomyocytes of said sample with more than one antibody or antigen binding fragment thereof specific for antigens of non-cardiomyocytes and c) depleting said labeled non-cardiomyocytes from said sample; and optionally d) labeling the cardiomyocytes of said sample with at least one antibody or antigen binding fragment thereof specific for antigen(s) of cardiomyocytes; and e) enriching said labeled cardiomyocytes and detecting and isolating cardiomyocyte subtypes derived from said pluripotent and/or multipotent stem cells.

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Bergisch Gladbach, Germany

Kristin Becker