Multi-layer airway organoids and methods of making and using the same
AssigneeWake Forest University Health Sciences
InventorsSean V. Murphy, Anthony Atala
Life of this patent
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Abstract
Provided herein are artificial lung organoids. The artificial lung organoids may include an epithelial cell layer comprising mammalian lung epithelial cells, a stromal cell layer comprising mammalian lung fibroblast cells and an endothelial cell layer comprising mammalian endothelial cells. The artificial lung organoids may optionally include a porous membrane between said epithelial cell layer and said stromal cell layer and/or between said stromal cell layer and said endothelial lung cell layer.
Claims20
A method of making a lung organoid, the method comprising:
- depositing an endothelial cell layer comprising mammalian endothelial cells onto a first side of a porous membrane;
- depositing a stromal cell layer comprising mammalian lung fibroblast cells onto a second side of the porous membrane that is opposite the first side of the porous membrane;
- and
- depositing an epithelial cell layer comprising human lung epithelial cells directly onto the stromal cell layer,
- thereby making the lung organoid, wherein cells of the epithelial cell layer of the lung organoid express at least one marker selected from the group consisting of p63+ KRT5+, Foxj1+ Sox17+, Scgb1A1+, Muc5ac+, and CFTR+.
A method of making a lung organoid, the method comprising:
- providing a porous membrane;
- coating a first side of the porous membrane and a second side that is opposite the first side of the porous membrane with an extracellular matrix composition to provide a coated porous membrane;
- depositing an endothelial cell layer comprising mammalian endothelial cells onto the first side of the coated porous membrane;
- depositing a stromal cell layer comprising lung fibroblast cells onto the second side of the coated porous membrane;
- and
- depositing an epithelial cell layer comprising human lung epithelial cells directly onto the stromal cell layer,
- thereby making the lung organoid, wherein cells of the epithelial cell layer of the lung organoid express at least one marker selected from the group consisting of p63+ KRT5+, Foxj1+ Sox17+, Scgb1A1+, Muc5ac+, and CFTR+.
Dependencies are inferred from the claim text.