Posters

Can limbal derived proteins potentiate transdifferentiation of dental pulp stem cells into a corneal epithelium for potential treatment of limbal stem cell deficiency?

Poster Details

First Author: L.Tanner UK

Co Author(s):    C. Hillarby   F. Carley   J. Yates   E. Kushnerev           

Abstract Details

Purpose:

Ocular surface damage secondary to chemical injury can permanently hinder the limbal stem cells ability to maintain normal corneal epithelial homeostasis, essential for good vision. The purpose of this study is to transdifferentiate dental pulp stem cells (DPSCs) with two abundant proteins found in the extracellular matrix of the limbus, keratocyte growth factor (KGF) and collagen type IV (Col4) into a corneal-like phenotype. DPSCs were used as safe autologous source of stem cells. Following this, the study will evaluate the transdifferentiated cell’s ability to re-epithelialize a limbal stem cell deficiency ex-vivo model of an acid burned human cornea.

Setting:

Blond McIndoe Laboratory, University of Manchester, Faculty of Biological, Medical and health Sciences

Methods:

The Limbal Stem Cell Deficiency (LSCD) model was established in human corneas (n=12) using 5M HCL acid. Sorted dental pulp stem cells were cultured with KGF and Col4 for 5 days. Cells were analysed with immunocytochemistry to evaluate whether the cells had transdifferentiated by fluorescent staining with cytokeratin 3 and 12, markers of corneal epithelium. These cells were seeded, labelled and cultured on contact lenses. The treated acid burned corneas were frozen and cryosectioned. Samples were analysed for cytokeratin 3/12, cytokeratin 13/19 conjunctival markers and CD90, a stem cell marker.

Results:

Cells cultured with KGF and Col4 showed expression of corneal epithelium markers (CK3/12), evidencing their transdifferentiation. These cells also exhibited corneal epithelial-like morphology in culture. Corneal samples showed evidence of dental pulp stem cells on the anterior surface, expressing corneal epithelial markers (CK3/12) with conjunctival markers absent (CK13/19).

Conclusions:

Transdifferentiated dental pulp stem cells using KGF and Col4 can be successfully used to repair the ocular surface and re-epithelialize an ex-vivo acid burned human cornea. The results demonstrate that this process could be a non-surgical treatment for LSCD due to chemical burn.

Financial Disclosure:

None