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US 11,021,688 B2Granted
Derivation of human skin organoids from pluripotent stem cells
AssigneeIndiana University Research Annd Technology Corporation
InventorsKarl R. Koehler, Jiyoon Lee
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Abstract
Provided herein are methods for directing differentiation of human pluripotent stem cells into a three-dimensional multilayered skin composition comprising an epidermal layer, a dermal layer, and a plurality of cells capable of forming a functional hair follicle. Also provided herein are three-dimensional, multilayered engineered skin compositions and methods of using the same for drug screening, for screening compounds for effects on hair growth, and for other applications.
Claims15
A method of obtaining a three-dimensional multilayered human skin composition, the method comprising:
- (a) culturing human pluripotent stem cell aggregates in a chemically defined culture medium comprising FGF-2 and an inhibitor of Transforming Growth Factor Beta (TGFβ) mediated signaling for about 4 days, whereby non-neural epithelium forms within the aggregates;
- (b) treating the stem cell aggregates of (a) with FGF-2 and an inhibitor of Bone Morphogenetic Protein (BMP) for about 4 to about 8 days;
- (c) embedding the cultured aggregates of (b) in a porous substrate comprising extracellular matrix components;
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- (d) culturing the embedded aggregates of (c) in floating culture for at least 48 days under conditions that promote self-assembly of cells within the embedded aggregates into a three-dimensional, multilayered human skin composition comprising an epidermal layer, a dermal layer, and a plurality of cells capable of forming a functional hair follicle.
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