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

Graefe's Archive for Clinical and Experimental Ophthalmology(2016)

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摘要
Purpose To examine the effects of media and deswelling agents on human corneal endothelial and epithelial cell viability using a previously developed screening system. Methods 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 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-1® and propidium iodide) and intracellular reactive oxygen species (ROS). Results 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. Conclusions The use of serum-supplemented MEM-based media for corneal organ culture should be discontinued in favour of serum-free media like SFM.
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关键词
Corneal organ cultivation,Serum-free medium,Deswelling substance,Cell survival,Human corneal endothelium,Human corneal epithelium
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