title: Preservation of Ocular Epithelial Limbal Stem Cells: The New Frontier in Regenerative Medicine.
abstract: Significant advances have been made in the field of ocular regenerative medicine. Promising stem cell-based therapeutic strategies have been translated into the clinical practice over the last few decades. These new stem cell-based therapies offer the possibility of permanently restoring corneal epithelium in patients with severe disabling and blinding ocular surface disease. The European Union has already classified stem cell-based therapies as "medicinal products". Therefore, manipulation is strictly regulated according to the defined conditions of good manufacturing practice, with the production of stem cell therapeutics at only accredited production sites authorized by the national regulatory agencies. In this regard, as first medical products are licensed for commercial use in Europe enabling a more widespread access to a stem cell-based therapy, the need for safe, validated and reproducible techniques for ex vivo cultured tissue preservation and distribution are coming to the forefront of research. However, these provide various new challenges for biobanking industry such as the retention of viability, good functionality of stem cells and sterility issues. This chapter provides an overview of the current advances in the field of corneal/limbal epithelial stem cell culture preservation techniques using either hypothermic storage or cryopreservation methods, that were used in different culturing steps (from stem cell isolation to the ex vivo epithelial graft preparation), with the reported impact on the post-thawing product recovery.
authors.affiliation: Eye Hospital, University Medical Centre, Ljubljana, Slovenia.
authors.forename: Zala
authors.initials: Z
authors.lastname: Lužnik
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venice, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Bertolin
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venice, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Breda
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venice, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Ferrari
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venice, Italy.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Barbaro
authors.affiliation: Eye Hospital, University Medical Centre, Ljubljana, Slovenia.
authors.forename: Petra
authors.initials: P
authors.lastname: Schollmayer
authors.affiliation: Fondazione Banca degli Occhi del Veneto, Venice, Italy. stefano.ferrari@fbov.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferrari
mesh_terms: D018070:Biological Specimen Banks / Q000331:legislation & jurisprudence; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D005060:Europe; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D044968:Regenerative Medicine / Q000331:legislation & jurisprudence / Q000379:methods*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D054457:Tissue Scaffolds; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
keywords: Cell-based therapy; Corneal disease; Corneal epithelium; Cryopreservation; Culture methods; Hypothermic storage; Ocular surface; Regenerative medicine; Stem cells; Vitrification