Effect of Growth Conditions on the Transport of Gly-Sar across Rabbit Corneal Epithelial Cells (SIRC) and Retinal Pigmental Epithelial Cells (ARPE-19)

Abstract

Purpose: : To screen the effect of several factors (hydrocortisone, insulin-transferrin-selenium mix, collagen-fibronectin-laminin coating, and air water interface) on three different response factors (TEER, mannitol transport and Gly-Sar transport) in Rabbit Corneal Epithelial Cells (SIRC) and Retinal Pigmental Epithelial Cells (ARPE-19) cells using a two level factorial design.

Methods: : ARPE-19 and SIRC cells were grown on 12 well transwell^® membranes in the presence and absence of various factor combinations as generated in a two level, 4 factor, 3 response factorial design. The factorial was generated using statgraphics^TM software. The TEER was measured. The [¹⁴C] Mannitol and [³H] Gly-Sar transport were estimated using Beckman scintillation counter. The significance of the response variables was calculated using statgraphics^TM software.

Results: : Out of all the factors studied, the combination of four of them gave the maximum TEER value. The presence air on the apical side of the cell membrane in case of SIRC cells increased the TEER value and decreased the mannitol transport to some extent. The presence of fibronectin and laminin along with collagen instead of just collagen also played a role in increasing tight junctions. These factors did not appreciably affect the transport of Gly-Sar across both the cell lines. There was a slight effect on the TEER values of ARPE-19 cells also in the presence of air interface. Further optimization studies would be carried out to find the optimum concentrations of these factors.

Keywords: cornea: epithelium • retinal pigment epithelium