Cultivation of Human Corneal Endothelial Cells on a Pericellular Matrix Prepared from Human Decidua-Derived Mesenchymal Cells

Abstract

Purpose: : To investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM) as an animal-free substrate for a human corneal endothelial cell (HCEC) culture for future clinical application.

Methods: : Decidua-derived mesenchymal cells (DMCs) were cultured on gelatin coated dishes for up to 3 days after they had reached confluence and were lysed by a deoxycholate treatment to prepare the PCM-DM. HCECs (on passage 3, the pre-experimental cell density was 1835 cells/mm²) were subcultured on a PCM-DM culture dish and a non-coated culture dish at a density of 5.0 x 10⁴/cm². Cell density and morphology were then evaluated for up to 2 months and the expression of function-related proteins such as ZO-1 and Na⁺/K⁺-ATPase was examined by immunocytochemistry.

Results: : The cell density of HCECs was found to be much higher on the PCM-DM culture dish compared to that on the non-coated culture dish (1803.0±25.4 and 1231.5±19.2 cells /mm², respectively; p<0.01). Moreover, a confluent monolayer of polygonal HCECs showing a homogeneous expression of ZO-1 and Na⁺/K⁺-ATPase were obtained on the PCM-DM culture dish.

Conclusions: : The findings of this present study indicate that PCM-DM can be effectively used as an animal-free substrate for culturing HCECs and that it enables a higher cell density to be maintained and the expression of function-related proteins in culture.

Keywords: cornea: endothelium • cornea: basic science • transplantation