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Quality assessment of corneal storage media and their components

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Abstract

Background

To keep the loss of endothelial cell density in donor corneas to a minimum, a storage medium which is adjusted to their nutritional needs is necessary. Different media, used either serum-supplemented or serum-free, are available. The quality of medium- and serum-batches as well as support of endothelial cell viability by the medium are to be tested with a quality assured screening system that allows routine examination.

Methods

A screening system was developed which is based on cell-culture tests with the well-established human corneal endothelial cell line HCEC-12, and therefore can be performed without the need for donor corneas. The cells are plated at a defined density in cell-culture dishes, and are cultured for a defined period of time in the test media. Evaluation is carried out by assaying cell count, activity of cell metabolism (resazurin conversion), and determining the number of apoptotic and necrotic cells (combined vital staining with YO-PRO®-1/propidium iodide and subsequent flow cytometry).

Results

Human corneal endothelial cells that are cultured in a medium which is adjusted to their nutritional needs achieve higher cell numbers and show a higher metabolic rate. Simultaneously, the percentage of apoptotic and necrotic cells is lower. The screening system developed in this study allows for easy and reliable detection of slightest differences between different media, different processing steps for same media, and different supplements, as well as different serum batches.

Conclusions

The differentiated results show that the screening system is sensitive enough to show even minor quality differences. Therefore, it is more suitable than the hitherto commonly used growth assay with primary, mostly porcine, corneal endothelial cells.

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Acknowledgments

This work was partially supported by DGFG (German Society for Tissue Transplantation, Hannover, Germany).

Conflict of interest

None

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Authors and Affiliations

  1. Coating of Flat Substrates, Medicine–Technical Application, Fraunhofer Institute for Electron Beam and Plasma Technology FEP, Winterbergstrasse 28, 01277, Dresden, Germany

    Jessy Schönfelder & Christiane Wetzel

  2. Institute of Anatomy, Faculty of Medicine Carl Gustav Carus, TU Dresden, Germany

    Monika Valtink, Lilla Knels, Richard H. W. Funk & Katrin Engelmann

  3. CRTD/DFG Center for Regenerative Therapies Dresden — Cluster of Excellence, TU Dresden, Dresden, Germany

    Richard H. W. Funk & Katrin Engelmann

  4. Department of Ophthalmology, Klinikum Chemnitz gGmbH, Chemnitz, Germany

    Katrin Engelmann

Authors
  1. Jessy Schönfelder
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  2. Monika Valtink
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  3. Lilla Knels
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  4. Richard H. W. Funk
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  5. Katrin Engelmann
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  6. Christiane Wetzel
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Corresponding author

Correspondence to Jessy Schönfelder.

Additional information

Presentation at a conference: 110. DOG-Kongress, September 20–23 2012, Berlin

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Schönfelder, J., Valtink, M., Knels, L. et al. Quality assessment of corneal storage media and their components. Graefes Arch Clin Exp Ophthalmol 252, 77–82 (2014). https://doi.org/10.1007/s00417-013-2482-5

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  • DOI: https://doi.org/10.1007/s00417-013-2482-5

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