Microfabricated niches for the culture of limbal epithelial cells

Abstract

Purpose: To explore whether the inclusion of micropockets or microfabricated niches mimicking the limbal stem cell niche provide any protection to cultured limbal cells when designing a biodegradable alternative to the amniotic membrane for limbal stem cell transplantation.

Methods: Micropockets in synthetic, degradable, poly-lactide-co-glycolide (PLGA) membranes were created by comb

Results: The presence of micropockets increased cell migration by 2-fold when compared to that on plain membranes under static conditions and also when subjected to flow stress. Positive staining for p63 and p63α showed the presence of limbal stem cells in membranes with and without micropockets. Positive staining for cytokeratin 3/12, a corneal epithelium specific marker, further confirmed the epithelial phenotype of the cultured cells.

Conclusions: The results of this study show that subjecting limbal epithelial cells to flow stress did not alter their epithelial phenotype. Further, the inclusion of microfabricated pockets in scaffolds resulted in higher cell density and greater cell migration under both static and dynamic culture conditions. Thus we suggest that the addition of micropockets to the synthetic membranes provides some physical advantage to the limbal epithelial cells over plain scaffolds.

Keywords: 482 cornea: epithelium • 721 stem cells • 741 transplantation