title: In vitro expansion and characterization of corneal stem cells isolated from an eye with malignant melanoma.
abstract: OBJECTIVE
  The objective of this study was the identification, characterization and in vitro replication of the human corneal stem cells, taking into consideration the difficulties in obtaining sufficient corneal material from living donors. The study explored a variety of stem cell markers, usually found in embryonic or adult mesenchymal stem cells. Culture medium and replication substrates had to be identified, with no data available on this subject in our country (there are no other reports on corneal stem cells in Romania, to our knowledge).
  
  
  METHODS
  Corneal epithelial limbus was harvested from an enucleated eye, containing also a choroid malignant melanoma. Stem cells from the limbus were isolated and cultivated in vitro. Expression of specific stem cell markers was evaluated with immunocytochemistry.
  
  
  RESULTS
  Corneal stem cell expansion in primary culture was slow, achieving 70-80% confluence after 28 days. Stem cells were easily isolated in standard medium, showed fibroblastoid morphology and were positive for certain stem cell specific markers in immunocytochemical staining: Oct3÷4, SOX2, Nanog, SSEA4, CD44, CD90, CD133, and CD34. They also expressed pan-cytokeratin. Donor age (72 years) and the presence of a malignant tumor close to limbal stem niche could have had an impact on the proliferation rate and the characteristics of the corneal stem cells.
  
  
  CONCLUSIONS
  Isolated limbal cells were adult type stem cells with an epithelial orientation. The characterization of these cells with immunocytochemistry allowed us to observe surface markers that other stem cells also express.
authors.affiliation: Ophthalmology Clinic, "Iuliu Hatieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania. iovidius@yahoo.com
authors.forename: O
authors.initials: O
authors.lastname: Samoilă
authors.forename: Olga
authors.initials: O
authors.lastname: Soriţău
authors.forename: M
authors.initials: M
authors.lastname: Călugăru
authors.forename: Lăcrămioara
authors.initials: L
authors.lastname: Totu
authors.forename: S
authors.initials: S
authors.lastname: Şuşman
authors.forename: Cristina
authors.initials: C
authors.lastname: Cristian
authors.forename: Carmen Mihaela
authors.initials: CM
authors.lastname: Mihu
mesh_terms: D000368:Aged; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology* / Q000473:pathology; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000473:pathology*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology