title: Improved preparation and preservation of human keratoplasty lenticules.
abstract: OBJECTIVE
  To improve the preparation of lenticules from human cornea and to obtain their preservation without loss of viable keratocytes.
  
  
  METHODS
  The epithelium was manually removed after bathing the surface of the cornea with a solution of trypsin and EDTA. Lenticules were prepared by microkeratome resection and viable keratocytes were visualized by staining with thiazolyl blue (MTT).
  
  
  RESULTS
  The pretreatment with trypsin-EDTA allowed the removal of the epithelium without damage to the keratocytes and the stroma. When these lenticules were incubated in Optisol-GS for 7 days at 4 degrees C, they showed a limited thickness increase and a preservation of keratocyte viability.
  
  
  CONCLUSIONS
  This procedure allows the preparation of lenticules with viable keratocytes that can be preserved in the cold for at least 1 week.
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venezia Mestre, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Salvalaio
authors.forename: Adriano
authors.initials: A
authors.lastname: Fasolo
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bruni
authors.forename: Anna Chiara
authors.initials: AC
authors.lastname: Frigo
authors.forename: Elisa
authors.initials: E
authors.lastname: Favaro
authors.forename: Diego
authors.initials: D
authors.lastname: Ponzin
mesh_terms: D002470:Cell Survival / Q000502:physiology; D002809:Chondroitin Sulfates; D045424:Complex Mixtures; D003315:Cornea; D003319:Corneal Stroma / Q000166:cytology*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000627:therapeutic use; D016895:Culture Media, Serum-Free; D003646:Debridement; D003911:Dextrans; D005347:Fibroblasts / Q000166:cytology*; D005839:Gentamicins; D006801:Humans; D015948:Keratoplasty, Penetrating; D009926:Organ Preservation / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D045424:Complex Mixtures; D003451:Cryoprotective Agents; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005839:Gentamicins; D002809:Chondroitin Sulfates; C075736:Optisol