title: Autologous fibrin-cultured limbal stem cells permanently restore the corneal surface of patients with total limbal stem cell deficiency.
abstract: BACKGROUND
  Ocular burns cause depletion of limbal stem cells, which leads to corneal opacification and visual loss. Autologous cultured epithelial cells can restore damaged corneas, but this technology is still developing. We sought to establish a culture system that allows preservation of limbal stem cells and preparation of manageable epithelial sheets and to investigate whether such cultures can permanently restore total limbal stem cell deficiency.
  
  
  METHODS
  We selected a homogeneous group of patients whose limbal cell deficiency was evaluated by scoring the gravity of the clinical picture and the keratin expression pattern. Stem cells, obtained from the limbus of the contralateral eye, were cultivated onto a fibrin substrate and their preservation was evaluated by clonal analysis. Fibrin cultures were grafted onto damaged corneas.
  
  
  RESULTS
  Fibrin-cultured limbal stem cells were successful in 14 of 18 patients. Re-epithelialization occurred within the first week. Inflammation and vascularization regressed within the first 3-4 weeks. By the first month, the corneal surface was covered by a transparent, normal-looking epithelium. At 12-27 months follow-up, corneal surfaces were clinically and cytologically stable. Three patients had a penetrating keratoplasty approximately 1 year after restoration of their corneal surface. Their visual acuity improved from light perception or counting fingers to 0.8-1.0.
  
  
  CONCLUSIONS
  Preservation of limbal stem cells in culture gives new perspectives on the treatment of ocular disorders characterized by complete limbal stem cell deficiency. The multicenter nature of this study and the handiness and ease of long-distance transportation of the fibrin-cultured epithelial sheets suggest that this technology can now be widely applied.
authors.affiliation: Division of Ophthalmology, Ospedale S.S. Giovanni e Paolo, Venice, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Rama
authors.forename: S
authors.initials: S
authors.lastname: Bonini
authors.forename: A
authors.initials: A
authors.lastname: Lambiase
authors.forename: O
authors.initials: O
authors.lastname: Golisano
authors.forename: P
authors.initials: P
authors.lastname: Paterna
authors.forename: M
authors.initials: M
authors.lastname: De Luca
authors.forename: G
authors.initials: G
authors.lastname: Pellegrini
mesh_terms: D016475:3T3 Cells; D000328:Adult; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003315:Cornea / Q000473:pathology; D016039:Corneal Transplantation / Q000502:physiology*; D005126:Eye Burns / Q000473:pathology / Q000628:therapy*; D005260:Female; D005337:Fibrin; D007839:Functional Laterality; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D016850:Limbus Corneae; D008297:Male; D051379:Mice; D008875:Middle Aged; D014182:Transplantation, Autologous
chemical_list: D005337:Fibrin