title: Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty.
abstract: To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts.
  
  
  
  50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK grafts were stored using three methods: peeling with free scroll storage; partial peeling with storage on the stroma and fluid bubble separation with storage on the stroma. Endothelial cell (EC) phenotype and the extent of endothelial overgrowth were examined. Global cell viability was assessed for storage methods that maintained a normal cell phenotype.
  
  
  
  1 mm wounds healed within 4 days. Enhanced media did not increase EC proliferation but may have increased EC migration into the wounded area. Grafts that had been trephined showed evidence of EC overgrowth, whereas preservation of a physical barrier in the bubble group prevented this. In grafts stored in enhanced media or reapposed to the stroma after trephination, endothelial migration occurred sooner and cells underwent endothelial-mesenchymal transformation. Ongoing cell loss, with new patterns of cell death, was observed after returning grafts to storage. Grafts stored as free scrolls retained more viable ECs than grafts prepared with the fluid bubble method (74.2± 3% vs 60.3±6%, p=0.04 (n=8).
  
  
  
  Free scroll storage is superior to liquid bubble and partial peeling techniques. Free scrolls only showed overgrowth of ECs after 4 days in organ culture, indicating a viable time window for the clinical use of pre-prepared DMEK donor material using this method. Methods for tissue preparation and storage media developed for whole corneas should not be used in pre-prepared DMEK grafts without prior evaluation.
authors.affiliation: Department of Corneal and External Disease, Moorfields Eye Hospital, London, UK.|University College London, Institute of Ophthalmology, London, UK.
authors.forename: Maninder
authors.initials: M
authors.lastname: Bhogal
authors.affiliation: University College London, Institute of Ophthalmology, London, UK.
authors.forename: Karl
authors.initials: K
authors.lastname: Matter
authors.affiliation: University College London, Institute of Ophthalmology, London, UK.
authors.forename: Maria S
authors.initials: MS
authors.lastname: Balda
authors.affiliation: Department of Corneal and External Disease, Moorfields Eye Hospital, London, UK.|University College London, Institute of Ophthalmology, London, UK.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Allan
mesh_terms: D000368:Aged; D002452:Cell Count; D002470:Cell Survival; D003316:Corneal Diseases / Q000601:surgery*; D057111:Descemet Stripping Endothelial Keratoplasty / Q000379:methods*; D004728:Endothelium, Corneal / Q000166:cytology*; D005260:Female; D006801:Humans; D008297:Male; D009924:Organ Culture Techniques; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods*; D014792:Visual Acuity
keywords: Cornea; Eye (Tissue) Banking; Imaging; Treatment Surgery; Wound healing