Patient-Specific iPSC-Derived Cornea Organoids for Investigating Aniridia-Associated Corneal Disorders

Aniridia is a rare congenital eye disorder marked by the partial or complete absence of the iris, and it is often associated with corneal opacification and limbal stem cell deficiency. The underlying genetic mutations, primarily affecting the PAX6 gene, disrupt both the development and function of the cornea. The most common ocular features besides iris defects are nystagmus, foveal hypoplasia, cataract, glaucoma and aniridia-associated keratopathy (AAK). Although in vivo models offer valuable insights into the pathology of aniridia, patient-derived in vitro models are crucial for investigating disease mechanisms and evaluating potential therapies. In this study, we present an in vitro AAK model using patient-derived induced pluripotent stem cells (iPSCs). Our protocol involves the stepwise differentiation of iPSCs into corneal epithelial-like cells within self-assembled three-dimensional organoids, which mimic the native corneal microenvironment. Through the optimization of differentiation culture conditions, we successfully generated corneal organoids that harbor distinct corneal cell populations, recapitulating the key structural and functional attributes of the human cornea.