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The existence of dead cells in donor corneal endothelium preserved with storage media
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  • Philippe Gain, Zhiguo He, Thibaud Garcin and Gilles Thuret
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    • Philippe Gain, Ophthalmic surgeon and researcher, head of the ophthalmology dept Laboratory "Biology, engineering and imaging of the Corneal graft", EA2521, University of St-Etienne, France
    • Other Contributors:
      • Zhiguo He, Research Engineer
      • Thibaud Garcin, Resident in ophthalmology
      • Gilles Thuret, Ophthalmic surgeon and researcher

    We noticed the article entitled “The existence of dead cells in donor corneal endothelium preserved with storage media” by Kitazawa with interest (Br J Ophthalmol 2017. Oct 5).
    Authors clearly demonstrated, using a triple staining with Hoechst, Propidium Iodide and Calcein-AM, that corneas stored at 4°C in Optisol-GS for 3 to 7 days, the technic most used worldwide, bear a significant number of dead endothelial cells (ECs). They underlined that these non-viable ECs are not recognized by specular cell count done by the eye bank and that this could explain the “cell loss” inevitably noticed post graft. Our team applied, for the first time in 2011 [1] and again in 2016 [2], a similar triple staining on the endothelium of whole corneas stored in long-term organ culture at 31°C, the dominant technic in Europe. We made the same findings as Kitazawa et al. and defined the notion of viable ECD (vECD) as the number of viable ECs per surface unit. Areas without ECs (especially in Descemetic folds), dead and dying EC (that will not survive the storage) clearly explain the important discrepancy between the cell count done by the eye bank (unable to spot them) and the very early postoperative ECD. Our team also demonstrated long ago that vital Trypan blue staining used by certain eye bank is unable to spot all dying cells because its time window of positivity is very narrow, corresponding only to ECs near to desquamate [3]. Viable ECD determined by triple staining therefore appea...

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    Conflict of Interest:
    None declared.