# High‐Purity Functional Corneal Endothelial Cells From Human Induced Pluripotent Stem Cells via a Novel Wash‐Out Method

## TL;DR

Scientists developed a new method to create high-purity corneal endothelial cells from stem cells, which could help treat blindness caused by corneal failure.

## Contribution

A novel wash-out method enables high-purity generation of functional corneal endothelial cells from human iPSCs.

## Key findings

- iPSC-derived CECs with wash-out show robust expression of CEC-specific markers and hexagonal morphology.
- Transplanted iPSC–CECs improved corneal transparency in a rabbit model without rejection.
- Single-cell sequencing confirmed iPSC–CECs are similar to primary human CECs.

## Abstract

Corneal endothelial failure can cause blindness, with transplantation as the only treatment. Due to donor shortages, establishing robust methods for generating corneal endothelial‐like cells (CECs) from induced pluripotent stem cells (iPSCs) is critical. Differentiation protocols included iPSC‐to‐CEC induction with or without neural crest cell differentiation. CECs directly differentiated from iPSCs demonstrated robust expression of CEC‐specific markers and a hexagonal morphology. The wash‐out protocol is a novel, efficient, noncytotoxic method for removing undifferentiated iPSCs and obtaining CEC populations with high purity. Single‐cell sequencing data showed that iPSC–CECs with wash‐out were similar to human primary CECs. In vivo transplantation of iPSC–CECs into a corneal endothelial dysfunction (CED) rabbit model demonstrated their safety and therapeutic efficacy, with improved corneal transparency. Notable recovery of corneal clarity in the CED model, without graft rejection, highlights the in vitro and in vivo potential of iPSC–CECs as a powerful source for clinical therapy in patients with CED. This work establishes an effective stem cell‐based platform for producing corneal endothelium‐like cells with clinical‐grade quality, offering a scalable and regenerative alternative to conventional transplantation.

iPSC‐derived corneal endothelial cells purified via a novel wash‐out method show high functional fidelity, resemble primary CECs, and restore corneal clarity in vivo, offering a scalable alternative to donor transplantation.