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Regenerative Medicine

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

    Graziella Pellegrini

    * Author for correspondence

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy.

    ,
    Paolo Rama

    San Raffaele Scientific Institute, Ophthalmology Unit, Milan, Italy

    Authors contributed equally

    Search for more papers by this author

    ,
    Stanislav Matuska

    San Raffaele Scientific Institute, Ophthalmology Unit, Milan, Italy

    ,
    Alessandro Lambiase

    Department of Ophthalmology, University of Rome Campus Biomedico, Rome, Italy

    ,
    Stefano Bonini

    Department of Ophthalmology, University of Rome Campus Biomedico, Rome, Italy

    ,
    Augusto Pocobelli

    Hospital San Giovanni Addolorata, Rome, Italy

    ,
    Rossella Gisoldi Colabelli

    Hospital San Giovanni Addolorata, Rome, Italy

    ,
    Leopoldo Spadea

    Department of Biotechnological & Applied Clinical Sciences, Eye Clinic, University of L’Aquila, L’Aquila, Italy

    ,
    Romina Fasciani

    Department of Ophthalmology, Policlinic ‘A. Gemelli’, Catholic University ‘Sacro Cuore’, Rome, Italy

    ,
    Emilio Balestrazzi

    Department of Ophthalmology, Policlinic ‘A. Gemelli’, Catholic University ‘Sacro Cuore’, Rome, Italy

    ,
    Paolo Vinciguerra

    Ophthalmology Unit, Istituto Clinico Humanitas, Rozzano, Milan, Italy

    ,
    Pietro Rosetta

    Ophthalmology Unit, Istituto Clinico Humanitas, Rozzano, Milan, Italy

    ,
    Achille Tortori

    Ophthalmology Unit, ‘Ospedale dei Pellegrini’, Naples, Italy

    ,
    Marco Nardi

    Department of Surgical, Medical & Molecular Pathology, University of Pisa, Pisa, Italy

    ,
    Giovanna Gabbriellini

    University Hospital, Pisa, Italy and Ophthalmology Unit, Di.N.O.G.M.I., University of Genova, IRCCS San Martino IST, Genova, Italy

    ,
    Carlo E Traverso

    Ophthalmology Unit, Di.N.O.G.M.I., University of Genova, IRCCS San Martino IST, Genova, Italy

    ,
    Claudio Macaluso

    Ophthalmology Unit, University of Parma, Parma, Italy

    ,
    Lorena Losi

    Department of Life Sciences, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Antonio Percesepe

    Department of Medical & Surgical Sciences, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Beatrice Venturi

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Francesca Corradini

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Athanasios Panaras

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Antonio Di Rocco

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy

    ,
    Paolo Guatelli

    Holostem Terapie Avanzate S.r.l., Modena, Italy

    &
    Michele De Luca

    Centre for Regenerative Medicine ‘Stefano Ferrari’, University of Modena & Reggio Emilia, Modena, Italy

    Published Online:https://doi.org/10.2217/rme.13.43

    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.

    Papers of special note have been highlighted as: ▪ of interest ▪▪ of considerable interest

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