Biological parameters determining the clinical outcome of autologous cultures of limbal stem cells

Graziella Pellegrini, Paolo Rama, Stanislav Matuska, Alessandro Lambiase, Stefano Bonini, Augusto Pocobelli, Rossella Gisoldi Colabelli, Leopoldo Spadea, Romina Fasciani, Emilio Balestrazzi, Paolo Vinciguerra, Pietro Rosetta, Achille Tortori, Marco Nardi, Giovanna Gabbriellini, Carlo E. Traverso, Claudio Macaluso, Lorena Losi, Antonio Percesepe, Beatrice VenturiFrancesca Corradini, Athanasios Panaras, Antonio Di Rocco, Paolo Guatelli, Michele De Luca

Research output: Contribution to journalArticle


Aim: 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. Conclusion: 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.

Original languageEnglish
Pages (from-to)553-567
Number of pages15
JournalRegenerative Medicine
Issue number5
Publication statusPublished - Sep 2013


  • cell therapy
  • cornea
  • epithelium
  • limbus
  • p63
  • regenerative medicine
  • stem cells

ASJC Scopus subject areas

  • Biomedical Engineering
  • Embryology

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    Pellegrini, G., Rama, P., Matuska, S., Lambiase, A., Bonini, S., Pocobelli, A., Colabelli, R. G., Spadea, L., Fasciani, R., Balestrazzi, E., Vinciguerra, P., Rosetta, P., Tortori, A., Nardi, M., Gabbriellini, G., Traverso, C. E., Macaluso, C., Losi, L., Percesepe, A., ... De Luca, M. (2013). Biological parameters determining the clinical outcome of autologous cultures of limbal stem cells. Regenerative Medicine, 8(5), 553-567.