Characterization of cultivated Human Oral Mucosal Epithelial Cells (HOMEs) in an autologous culture medium

Abstract

Purpose: The aim of this study is to culture human oral mucosal cells (HOMEs) for reconstruction of human ocular surface. In the present study we characterize two different cell populations expanded in medium supplemented with autologous serum.

Methods: Small biopsy of human oral mucosal epithelium was harvested and after enzymatic treatment cultured in medium supplemented with autologous serum for 3 weeks at 37 degrees Celsius with 5% CO2 in a humidified atmosphere and the medium was changed every 3-4 days. Two different cell populations were then isolated and analyzed by qRT-PCR, electron microscopy (EM) and immunohistochemistry.

Results: In our study, two isolated populations (Pa and Pb) were successfully proliferated in medium supplemented with autologous serum compared with fetal bovine serum (FBS) containing medium. The levels of markers associated with proliferation, stemness, cell junction, and functional ocular surface such as KI67, ABCG2, P63, SOX9, CK19, CK3, CX43, E-CAD, and OCLN were differentially expressed in these two Pa and Pb populations, but in the Pa population stemness and cell junction related markers were extremely higher expressed compared with the Pb population.

Conclusions: The preliminary results of this study show that expanded human oral mucosal cells (HOMEs) in our culture system generates a committed cell population that can potentially be used in reconstraction of human ocular surface.

Keywords: 721 stem cells