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Banking of Donor Tissues for Descemet Stripping Automated Endothelial Keratoplasty

Ruzza, Alessandro MS; Salvalaio, Gianni BSn; Bruni, Alessandro MD; Frigo, Anna Chiara MSc; Busin, Massimo MD; Ponzin, Diego MD

doi: 10.1097/ICO.0b013e31825e8442
Basic Investigation

Purpose: The demands for precut lamellar grafts for Descemet stripping automated endothelial keratoplasty rose in our eye bank from 74 in 2007 to 408 in 2010. To meet this expanding requirement, we explored the possibility to preserve these preparations in organ culture.

Methods: Organ cultured corneas, stored in a medium containing 6% dextran, were mounted on a Moria artificial anterior chamber, deprived of the epithelium and then cut with a microkeratome. The posterior lamella was protected by positioning the anterior stromal cap, trephined at a diameter of 8.5 mm and stored at 31°C in the medium containing dextran. The endothelium was examined with trypan blue and alizarin staining and tested for its glycolytic activity (conversion of glucose into lactate).

Results: Incubation for a period of 1 week caused a small increase in the cell loss observed after trephination (from 6.2% to 10.6%). After 2 weeks, the decrease in endothelial cell density was 19.9% but the endothelial organization remained intact. The rate of glycolysis remained unchanged during the 2 weeks of preservation, with the majority of glucose uptake accounted for by lactate production. The thickness of the lenticules remained constant, ranging from 170 to 180 μm during the preservation.

Conclusions: The lamellar grafts for Descemet stripping automated endothelial keratoplasty may be stored in organ culture for 2 weeks without damaging the endothelium or increasing the overall thickness.

*Fondazione Banca degli Occhi del Veneto, Venezia, Italy

Dipartimento di Scienze Cardiologiche, Toraciche e Vascolari, Università degli Studi di Padova, Padova, Italy

Ophthalmic Department, Villa Igea Hospital, Forlì, Italy.

Reprints: Diego Ponzin, Fondazione Banca degli Occhi del Veneto, Via Paccagnella 11, 30174 Zelarino, Venezia, Italy (e-mail:

M. Busin receives travel reimbursements and royalties from Moria (Antony, France).

The other authors declare no conflicts.

Received February 2, 2012

Accepted May 8, 2012

Copyright © 2013 Wolters Kluwer Health, Inc. All rights reserved.