Characterization of Biomaterial free Cell Sheets Cultured from Human Oral Mucosal Epithelial Cells

Abstract

Purpose: To investigate the characteristics of biomaterial free cell sheets cultured from oral mucosal epithelial cells both in vitro and in a limbal deficient animal model.

Methods: Both human oral mucosal epithelial cells and limbal epithelial cells were cultured to form cell sheets with or without fibrin glue for 2 weeks. The resulting sheets were detached using a buffer containing 1% dispase and transplanted to limbal deficient rabbits which had been chemically injured with lamellar limbectomy. The adhesion stability of these biomaterial-free cell sheets were evaluated three days after transplantation. In parallel, Colony Forming Efficiency(CFE) as well as the immunohistochemistry using antibodies specific to proliferation and differentiation of epithelial cells were performed to characterize the cell sheets.

Results: CFE in Human oral mucosal epithelial cells and limbal epithelial cells were measured to be 57.5% and 14%, respectively. K1, K3, and K4 were expressed in mucosal epithelial sheets while K3 and K12 were in limbal epithelial sheets. The cell sheets were composed of three to six layers of stratified, well differentiated mucosal epithelial cells with 87.5% viable cells present. The in vivo adhesion stability of biomaterial-free cell sheets of oral mucosal epithelial cells was comparable to that of fibrin-based counterpart.

Conclusions: Our results suggest that biomaterial free cell sheets of oral mucosal epithelial cells can be a good candidate in the treatment of limbal deficient diseases.

Keywords: 721 stem cells