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Graefes Arch Clin Exp Ophthalmol. 2016 Feb;254(2):285-95. doi: 10.1007/s00417-015-3235-4. Epub 2015 Dec 19.

Effect of different culture media and deswelling agents on survival of human corneal endothelial and epithelial cells in vitro.

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Institute of Anatomy, Faculty of Medicine Carl Gustav Carus of the TU Dresden, Fetscherstr. 74, 01307, Dresden, Germany.
Institute of Anatomy, Faculty of Medicine Carl Gustav Carus of the TU Dresden, Fetscherstr. 74, 01307, Dresden, Germany.
Department of Ophthalmology, Klinikum Chemnitz gGmbH, Flemmingstr. 2, 09116, Chemnitz, Germany.



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, CorneaMax®/CorneaJet®, serum-free: Human Endothelial-SFM, Stemalpha-2 and -3) with and without 6 % dextran T500 or 7 % HES 130/0.4. Standard growth media F99HCEC and DMEM/F12HCE-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-1® 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 CorneaMax®, CorneaJet®, and CorneaMax® 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 CorneaJet®, but not by CorneaMax® 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.


Cell survival; Corneal organ cultivation; Deswelling substance; Human corneal endothelium; Human corneal epithelium; Serum-free medium

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