title: Biological parameters determining the clinical outcome of autologous cultures of limbal stem cells.
abstract: OBJECTIVE
  Limbal cultures restore the corneal epithelium in patients with ocular burns. We investigated the biological parameters instrumental for their clinical success.
  
  
  METHODS
  We report a long-term multicenter prospective study on 152 patients carrying corneal destruction due to severe ocular burns, treated with autologous limbal cells cultured on fibrin and clinical-grade 3T3-J2 feeder cells. Clinical results were statistically evaluated both by parametric and nonparametric methods.
  
  
  RESULTS
  Clinical outcomes were scored as full success, partial success and failure in 66.05, 19.14 and 14.81% of eyes, respectively. The total number of clonogenic cells, colony size, growth rate and presence of conjunctival cells could not predict clinical results. Instead, the clinical data provided conclusive evidence that graft quality and likelihood of a successful outcome rely on an accurate evaluation of the number of stem cells detected before transplantation as holoclones expressing high levels of the p63 transcription factor. No adverse effects related to the feeder layer have been observed and the regenerated epithelium was completely devoid of any 3T3-J2 contamination.
  
  
  CONCLUSIONS
  Cultures of limbal stem cells can be safely used to successfully treat massive destruction of the human cornea. We emphasize the importance of a discipline for defining the suitability and the quality of cultured epithelial grafts, which are relevant to the future clinical use of any cultured cell type.
authors.affiliation: Centre for Regenerative Medicine 'Stefano Ferrari', University of Modena & Reggio Emilia, Modena, Italy. graziella.pellegrini@unimore.it
authors.forename: Graziella
authors.initials: G
authors.lastname: Pellegrini
authors.forename: Paolo
authors.initials: P
authors.lastname: Rama
authors.forename: Stanislav
authors.initials: S
authors.lastname: Matuska
authors.forename: Alessandro
authors.initials: A
authors.lastname: Lambiase
authors.forename: Stefano
authors.initials: S
authors.lastname: Bonini
authors.forename: Augusto
authors.initials: A
authors.lastname: Pocobelli
authors.forename: Rossella Gisoldi
authors.initials: RG
authors.lastname: Colabelli
authors.forename: Leopoldo
authors.initials: L
authors.lastname: Spadea
authors.forename: Romina
authors.initials: R
authors.lastname: Fasciani
authors.forename: Emilio
authors.initials: E
authors.lastname: Balestrazzi
authors.forename: Paolo
authors.initials: P
authors.lastname: Vinciguerra
authors.forename: Pietro
authors.initials: P
authors.lastname: Rosetta
authors.forename: Achille
authors.initials: A
authors.lastname: Tortori
authors.forename: Marco
authors.initials: M
authors.lastname: Nardi
authors.forename: Giovanna
authors.initials: G
authors.lastname: Gabbriellini
authors.forename: Carlo E
authors.initials: CE
authors.lastname: Traverso
authors.forename: Claudio
authors.initials: C
authors.lastname: Macaluso
authors.forename: Lorena
authors.initials: L
authors.lastname: Losi
authors.forename: Antonio
authors.initials: A
authors.lastname: Percesepe
authors.forename: Beatrice
authors.initials: B
authors.lastname: Venturi
authors.forename: Francesca
authors.initials: F
authors.lastname: Corradini
authors.forename: Athanasios
authors.initials: A
authors.lastname: Panaras
authors.forename: Antonio
authors.initials: A
authors.lastname: Di Rocco
authors.forename: Paolo
authors.initials: P
authors.lastname: Guatelli
authors.forename: Michele
authors.initials: M
authors.lastname: De Luca
mesh_terms: D002452:Cell Count; D049109:Cell Proliferation; D002478:Cells, Cultured; D002999:Clone Cells; D003114:Colony-Forming Units Assay; D019573:Epithelium, Corneal / Q000637:transplantation; D005260:Female; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D008297:Male; D008875:Middle Aged; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology*; D014182:Transplantation, Autologous; D016896:Treatment Outcome; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D014792:Visual Acuity
chemical_list: D025521:Tumor Suppressor Proteins