High-Purity Functional Corneal Endothelial Cells From Human Induced Pluripotent Stem Cells via a Novel Wash-Out...

Area of Science:

• Ophthalmology
• Regenerative Medicine
• Stem Cell Biology

Background:

• Corneal endothelial failure leads to blindness, with limited donor availability for transplantation.
• Induced pluripotent stem cells (iPSCs) offer a potential source for generating corneal endothelial-like cells (CECs).

Purpose of the Study:

• To develop and validate a robust method for generating clinical-grade CECs from iPSCs.
• To assess the safety and efficacy of iPSC-derived CECs in a preclinical model of corneal endothelial dysfunction.

Main Methods:

• iPSCs were differentiated into CECs using specific protocols, with and without neural crest cell induction.
• A novel wash-out technique was employed to purify CEC populations.
• Single-cell sequencing was used to compare iPSC-CECs with primary human CECs.
• In vivo transplantation of iPSC-CECs was performed in a rabbit model of corneal endothelial dysfunction.

Main Results:

• Direct iPSC-to-CEC differentiation yielded cells with characteristic CEC markers and morphology.
• The wash-out protocol efficiently removed undifferentiated iPSCs, achieving high CEC purity.
• iPSC-CECs closely resembled primary human CECs based on single-cell sequencing.
• Transplanted iPSC-CECs demonstrated safety and efficacy, improving corneal transparency in the rabbit model without rejection.

Conclusions:

• A scalable stem cell-based platform for producing clinical-grade iPSC-CECs has been established.
• iPSC-derived CECs represent a viable regenerative alternative to traditional corneal transplantation for treating corneal endothelial dysfunction.
• This approach holds significant potential for addressing donor shortages and treating corneal blindness.