Article Text

Download PDFPDF

P02-A121 Precut DSAEK stored in an active storage machine: feasibility study
  1. Sylvain Poinard1,2,
  2. Oliver Dorado1,2,
  3. Paul Goin1,3,
  4. Chantal Perrache1,
  5. Zhiguo He1,
  6. Sandrine Ninotta1,
  7. Frédéric Mascarelli1,4,
  8. Philippe Gain1,2,
  9. Gilles Thuret1,2,
  10. Anne-Sophie Gauthier1,3
  1. 1Laboratoire de Biologie, Ingénierie et Imagerie pour l’Ophthalmology, BiiO, Faculté de Médecine, Université de Jean Monnet, 10 rue de la Marandière, 42270 Saint-Etienne, France
  2. 2Département Ophtalmologie, Centre Hospitalier Universitaire, Avenue Albert Raimond, 42055 Saint-Etienne Cedex 02, France
  3. 3Département Ophtalmologie, Centre Hospitalier Universitaire, 25000 Besançon, France
  4. 4Centre de Recherche des Cordeliers, UMR S1138, Université Paris Descartes, Paris, France


Purpose The number of endothelial grafts precut by eye banks increases. Their shelf life is limited to a few days. We previously demonstrated the superiority of an active storage machine (ASM) over organ culture (passive) for whole corneas. Aims: to measure endothelial viability of precut DSAEK after 3 or 10 days of storage in our ASM in a preclinical study.

Methods Human pairs of corneas were included. The endothelial cell density (ECD in cells/mm2), and central corneal thickness (CCT in μm) were measured to ensure their initial intra pair comparability. After deswelling (CorneaJet, Eurobio) grafts preparation was performed by cutting the anterior stroma with a Moria linear microkeratome and keeping the anterior lamellae attached during storage. After randomization, one cornea was kept in the corneajet bottle (CJ) and the other was inserted into the ASM allowing a renewal or storage medium (CorneaMax, Eurobio) at 2.6 µL/min with 21 mmHg of pressure in the endothelial chamber. Both group of corneas were stored for 3 or 10 days at 31°C. The final viable ECD (vECD) was determined using the triple staining with Hoechst-Ethidium-Calcein-AM by an independent experimenter in a masked fashion.

Results Initial ECDs were comparable: 2595±878 in ASM versus 2654±954 cells/mm2 in CJ for the 3-days period (n=5 pairs) and 2416±712 in ASM versus 2492±764 cells/mm2 in CJ for the 10-period (n=5 pairs). CCTs were also comparable. The anterior lamellae stayed attached in either the ASM or CJ. vECD was significantly higher in ASM than in CJ with respectively 2062±695 cells/mm2 versus 1632±633 cells/mm2 after 3 days either a cell loss of 20.5% and 38.5% respectively (p=0.0062) and 1082±649 versus 935±691 cells/mm2 for the 10-day period either a cell loss of 132% and 164% respectively (p=0.005). Grafts thickness did not differ after 3 days 219±25 µm in ASM versus 182±39 µm (p=0.063) or 10 days respectively 221±58 µm versus 189±48 µm (p=0.06).

Conclusion The storage of precut DSAEKs into the ASM allows a better preservation of grafts without use on deswelling storage medium. Nevertheless, the cell loss remains high after 10 days, suggesting a significant cell stress.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See:

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.