To examine the effects of media and deswelling agents on human corneal endothelial and epithelial cell viability using a previously developed screening system.

The human corneal endothelial cell line HCEC-12 and the human corneal epithelial cell line HCE-T were cultured in four different corneal organ culture media (serum-supplemented: MEM +2 % FCS, CorneaMaxA (R)/CorneaJetA (R), serum-free: Human Endothelial-SFM, Stemalpha-2 and -3) with and without 6 % dextran T500 or 7 % HES 130/0.4. Standard growth media F99(HCEC) and DMEM/F12(HCE-T) served as controls. In additional controls, the stress inducers staurosporine or hydrogen peroxide were added. After 5 days in the test media, cell viability was assessed by flow cytometrically quantifying apoptotic and necrotic cells (sub-G1 DNA content, vital staining with YO-PRO-1A (R) and propidium iodide) and intracellular reactive oxygen species (ROS).

The MEM-based media were unable to support HCEC-12 and HCE-T survival under stress conditions, resulting in significantly increased numbers of apoptotic and necrotic cells. HCEC-12 survival was markedly improved in SFM-based media even under staurosporine or hydrogen peroxide. Likewise, HCE-T survival was improved in SFM with or without dextran. The media CorneaMaxA (R), CorneaJetA (R), and CorneaMaxA (R) with HES supported HCEC-12 survival better than MEM-based media, but less well than SFM-based media. HCE-T viability was also supported by CorneaJetA (R), but not by CorneaMaxA (R) with or without HES. Stemalpha-based media were not suitable for maintaining viability of HCEC-12 or HCE-T in the applied cell culture system.

The use of serum-supplemented MEM-based media for corneal organ culture should be discontinued in favour of serum-free media like SFM.