Rapidly constructed scaffold-free embryonic stem cell sheets for ocular surface reconstruction

Area of Science:

• Ophthalmology
• Stem Cell Biology
• Tissue Engineering

Background:

• Limbal stem cell deficiency (LSCD) impairs corneal regeneration.
• Previous work optimized embryonic stem cell (ESC) sheet generation via centrifugation.
• The therapeutic potential of ESC sheets for ocular surface reconstruction requires further investigation.

Purpose of the Study:

• To evaluate the efficacy of scaffold-free embryonic stem cell sheets (SESCS) for ocular surface reconstruction in a rabbit model of LSCD.
• To assess the differentiation and integration of transplanted ESCs.
• To investigate the potential of ESCs as a cell source for corneal tissue engineering.

Main Methods:

• LSCD was induced in rabbits.
• Treated group received SESCS; control group received no treatment.
• Ocular surface evaluation included slit lamp observation, immunofluorescence (Cytokeratin K3, MUC5AC, OCT-4), cell tracing (DiIC18(3)-DS), and Atomic Force Microscopy (AFM).

Main Results:

• SESCS facilitated ocular surface reconstruction in 75% of treated rabbits.
• Control group exhibited conjunctivalization and neovascularization.
• SESCS group showed positive K3, negative MUC5AC, and OCT-4 expression, indicating corneal epithelial differentiation.
• Labeled cells were detected in vivo up to 15 days post-transplant.
• AFM identified three distinct cell types in the regenerated corneal epithelium.

Conclusions:

• Scaffold-free ESC sheets are effective in reconstructing the ocular surface in LSCD rabbits.
• ESCs can differentiate into corneal epithelial cells upon stromal contact.
• SESCS represent a promising cell source for corneal tissue engineering applications.