title: A novel method for preservation of human corneal limbal tissue.
abstract: OBJECTIVE
  We investigated the efficacy of low-temperature airlift preservation of human corneal limbal tissue for ex vivo expansion and allograft keratolimbal transplantation.
  
  
  METHODS
  Human limbal tissue either was submerged or airlifted in Optisol-GS medium and preserved at 4°C for up to eight days. Hematoxylin and eosin, and E-cadherin staining was performed to investigate epithelial structure and cell-cell junction. Epithelial cell differentiation and proliferation were studied using the biomarkers, such as K10, K12, K14, Ki67, and p63. Cell apoptosis was detected with the TUNEL assay. The epithelial progenitor cell pool was evaluated by clonal culture of epithelial cells on 3T3 feeder layers. For clinical application, keratolimbal transplantation was performed in three patients with partial limbal stem cell deficiency, using limbal tissues preserved under the airlift manner. Pre- and postoperative evaluations were conducted by slit-lamp microscopy and fluorescein staining.
  
  
  RESULTS
  After eight days, intact epithelia with strong cell-cell junctions were retained only in airlifted tissues. Airlifting maintained a normal corneal differentiation pattern, along with low proliferation activity and increased proliferation potential, but little apoptosis. Epithelial cells harvested from the airlift preservation for up to eight days exhibited stable clonogenicity. Limbal tissues preserved under the airlift manner successfully reconstructed corneal and limbal surfaces in partial limbal stem cell-deficient patients.
  
  
  CONCLUSIONS
  Limbal tissues preserved under hypothermic airlift conditions maintain the intact structure, normal phenotype, high viability, and stem cell pool of limbal epithelia. Such a method may be used in eye bank tissue processing and corneal epithelial tissue engineering.
authors.affiliation: Eye Institute and Affiliated Xiamen Eye Center, Xiamen University School of Medicine, Xiamen, Fujian, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Li
authors.forename: Nuo
authors.initials: N
authors.lastname: Dong
authors.forename: Huping
authors.initials: H
authors.lastname: Wu
authors.forename: Fei
authors.initials: F
authors.lastname: Dong
authors.forename: Yajie
authors.initials: Y
authors.lastname: Xu
authors.forename: Huiyi
authors.initials: H
authors.lastname: Du
authors.forename: Hui
authors.initials: H
authors.lastname: He
authors.forename: Zuguo
authors.initials: Z
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Li
mesh_terms: D016475:3T3 Cells; D000328:Adult; D000369:Aged, 80 and over; D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002102:Cadaver; D002454:Cell Differentiation; D049109:Cell Proliferation; D002648:Child; D018920:Coculture Techniques; D003319:Corneal Stroma / Q000166:cytology / Q000378:metabolism; D016039:Corneal Transplantation / Q000379:methods*; D019573:Epithelium, Corneal / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D005126:Eye Burns / Q000601:surgery*; D006085:Graft Survival; D006801:Humans; D007365:Intercellular Junctions; D016850:Limbus Corneae / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D013234:Stem Cells / Q000166:cytology; D014019:Tissue Donors; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D015415:Biomarkers
keywords: cornea; limbus; preservation; stem cell