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

Ali Can Koç; Gaye Su Yiğit; Semih Numan Göçmez; Mustafa Titiz; Sinan Güven

doi:10.3791/68556

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Method Article

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

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DOI:

10.3791/68556

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July 28th, 2026

Ali Can Koç¹^,² , Gaye Su Yiğit¹^,² , Semih Numan Göçmez¹^,² , Mustafa Titiz¹^,² , Sinan Güven¹^,²^,³

¹Izmir Biomedicine and Genome Center, ²Izmir International Biomedicine and Genome Institute, Dokuz Eylül University, ³Department of Medical Biology and Genetics, Faculty of Medicine, Dokuz Eylül University

Summary

We describe a protocol for generating cornea organoids from iPSCs derived from aniridia patients, creating a patient-specific model to study corneal disorders. These organoids mimic key structural and functional features, providing a reliable in vitro platform for investigating disease mechanisms, testing potential treatments, and advancing personalized regenerative approaches for anterior eye diseases.

Abstract

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.

Introduction

The cornea is an avascular, transparent tissue forming the outermost layer of the eye, consisting mainly of three layers: an epithelial outer layer, a stromal layer containing a collagen-rich extracellular matrix populated by keratocytes, and an endothelial cell layer¹. The cornea plays an essential role in vision by protecting internal ocular structures and refracting light.

Aniridia is a rare, congenital panocular disorder primarily caused by heterozygous mutations in the paired box gene 6 (PAX6), a master regulator of ocular development, and is characterized by partial or complete absence of the ....