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

US12221630B2 · kind B2 · utility

Assignee

• MILTENYI BIOTEC B.V. & CO. KG · DE

Inventors

• David Joel AGORKU · Cologne, DE
• Olaf Hardt · Cologne, DE
• Andreas Bosio · Cologne, DE
• Kristin Becker · Moitzfeld, DE
• Dominik Eckardt · Moitzfeld, DE

Key dates

• Filing date: Feb 18, 2020
• Grant date: Feb 11, 2025
• Expiry date: Mar 4, 2042

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2533/70
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

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 a…