title: Donor cornea transfer from Optisol GS to organ culture storage: a two-step procedure to increase donor tissue lifespan.
abstract: OBJECTIVE
  Storage time for donor corneas in Optisol GS is limited compared to Eye Bank Organ Culture (EBOC). We here examine the epithelium on donor corneoscleral rims after primary storage in Optisol GS and subsequent incubation in EBOC.
  
  
  METHODS
  Morphology was monitored by light and electron microscopy, expression of phenotypic and genotypic markers by immunohistochemistry and RT-PCR and changes in oxidative lipid and DNA damage by ELISA and COMET assay.
  
  
  RESULTS
  A prominent loss of cells was observed after storage in Optisol GS. After maintenance in EBOC, spreading apical cells were Occludin(+) , while the staining for E-cadherin and Connexin-43 was less intense. There were an upregulation of Occludin and a downregulation of E-cadherin and Connexin-43. Eye Bank Organ Culture was associated with an ongoing proliferative activity and a downregulation of putative progenitor/stem cell marker ABCG2 and p63. Staining for 8-OHdG and Caspase-3 did not increase, while levels of malondialdehyde and number of DNA strand breaks and oxidized bases increased.
  
  
  CONCLUSIONS
  This dual procedure should be pursued as an option to increase the storage time and the pool of available donor corneas. The observed downregulation of markers associated with stemness during EBOC is relevant considering the potential use of donor epithelium in the treatment of ocular surface disorders.
authors.affiliation: Center for Eye Research, Department of Ophthalmology, Oslo University Hospital, Ullevål and University of Oslo, Oslo, Norway. kristi5@medisin.uio.no
authors.forename: Kristiane
authors.initials: K
authors.lastname: Haug
authors.forename: Amaya
authors.initials: A
authors.lastname: Azqueta
authors.forename: Siv
authors.initials: S
authors.lastname: Johnsen-Soriano
authors.forename: Aboulghassem
authors.initials: A
authors.lastname: Shahdadfar
authors.forename: Liv K
authors.initials: LK
authors.lastname: Drolsum
authors.forename: Morten C
authors.initials: MC
authors.lastname: Moe
authors.forename: Magnus T
authors.initials: MT
authors.lastname: Røger
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Romero
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Collins
authors.forename: Bjørn
authors.initials: B
authors.lastname: Nicolaissen
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation; D002809:Chondroitin Sulfates / Q000627:therapeutic use*; D020552:Comet Assay; D045424:Complex Mixtures / Q000627:therapeutic use; D003315:Cornea; D015925:Cryopreservation / Q000379:methods*; D016895:Culture Media, Serum-Free; D004249:DNA Damage; D003911:Dextrans / Q000627:therapeutic use*; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal / Q000378:metabolism / Q000648:ultrastructure; D005125:Eye Banks / Q000379:methods*; D005260:Female; D005839:Gentamicins / Q000627:therapeutic use*; D006801:Humans; D020287:In Situ Nick-End Labeling; D015227:Lipid Peroxidation; D008054:Lipid Peroxides / Q000235:genetics / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009924:Organ Culture Techniques; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000627:therapeutic use*; D018384:Oxidative Stress; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014019:Tissue Donors
chemical_list: D015415:Biomarkers; D045424:Complex Mixtures; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005839:Gentamicins; D008054:Lipid Peroxides; D019188:Organ Preservation Solutions; D002809:Chondroitin Sulfates; C075736:Optisol