US 12,741,061 B2 · Method for obtaining healthy intestinal organoids

Abstract

The present invention is related to a biofunctional three-dimensional hydrogel suitable for the expansion of freshly isolated or frozen intestinal cells and the formation of intestinal organoids therefrom, that avoids the use of a naturally-derived matrix such as Matrigel and provides intestinal organoids suitable for clinical applications and generated in a commercially feasible manner. The present invention is also related to a kit of parts comprising said hydrogel in combination with a suitable culture medium, and to a method of making organoids and expanding cells from freshly isolated or frozen intestinal cells with said kit of parts.

Claims15

Dependent claims

A biofunctional three-dimensional hydrogel suitable for the growth and the expansion of freshly isolated or frozen intestinal cells and the formation of intestinal organoids therefrom,

wherein said hydrogel is the reaction product of at least one precursor molecule, which is a multi-arm PEG containing ethylenically unsaturated groups selected from the group consisting of vinylsulfone and acrylate moieties,
a crosslinker molecule containing at least two nucleophilic groups capable of reacting with said ethylenically unsaturated groups of said multi-arm PEG in a Michael addition reaction, and
at least one biofunctional ligand selected from the group consisting of natural laminins and recombinant laminin isoforms, characterized in that the crosslinker molecule comprises at least one RGD motif.

Kit of parts for the growth and expansion of freshly isolated or frozen intestinal cells and the formation of intestinal organoids therefrom, comprising at least one precursor molecule, which is a multi-arm PEG containing ethylenically unsaturated groups selected from the group consisting of vinylsulfone and acrylate moieties,

a crosslinker molecule containing at least two nucleophilic groups capable of reacting with said ethylenically unsaturated groups of said multi-arm PEG in a Michael addition reaction,
wherein the crosslinker molecule comprises at least one RGD motif, and
at least one biofunctional ligand selected from the group consisting of natural laminins and recombinant laminin isoforms, and
a culture medium, wherein said culture medium comprises R-spondin 1 and Wnt3a.

1 more element

Dependencies are inferred from the claim text.

Method for obtaining healthy intestinal organoids

Life of this patent

Method for obtaining healthy intestinal organoids

Life of this patent

Abstract

Claims15