Derivation of human skin organoids from pluripotent stem cells

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.

1 .- 12 . (canceled) 
     
     
         13 . A three-dimensional, multilayered engineered skin composition comprising an epidermal layer comprising human pluripotent stem cell-derived epidermal keratinocytes, a dermal layer comprising human pluripotent stem cell-derived dermal fibroblasts, and a plurality of human pluripotent stem cell-derived cells capable of forming a functional hair follicle, wherein the epidermal layer and dermal layer are in direct contact. 
     
     
         14 . The engineered skin composition of  claim 13 , wherein the plurality of cells capable of forming a functional hair follicle comprises cells selected from the group consisting of mesenchymal stem cells, dermal papilla cells, dermal sheath cells, and follicular epidermal stem cells. 
     
     
         15 . The engineered skin composition of  claim 13 , further comprising at least one functional hair follicle, functional sebaceous gland, or sensory neuron, 
     
     
         16 . The engineered skin composition of  claim 13 , further comprising a scaffold. 
     
     
         17 . The engineered skin composition of  claim 16 , wherein the scaffold is biodegradable or bioabsorbable. 
     
     
         18 . The engineered skin composition of  claim 16 , wherein the scaffold is a synthetic matrix. 
     
     
         19 . The engineered skin composition of  claim 13 , wherein the composition is capable of viable transplantation and engraftment to a human subject. 
     
     
         20 . A method of testing a compound for effects on hair growth, the method comprising
 (a) contacting a test compound to a three-dimensional multilayered skin composition obtained according to the method of claim  1 ; and   (b) detecting an effect of the agent on one or more cell types within the contacted skin composition.   
     
     
         21 . The method of  claim 20 , wherein detecting comprises performing a method selected from the group consisting of RNA sequencing, gene expression profiling, transcriptome analysis, and protein expression analysis. 
     
     
         22 . The method of  claim 20 , wherein the agent is screened for an effect on gene expression and wherein detecting comprises assaying for differential gene expression relative to an uncontacted skin composition. 
     
     
         23 . Use of a three-dimensional multilayered skin composition obtained according to the method of claim  1  in a drug discovery screen.