title: Long-term phenotypic study after allogeneic cultivated corneal limbal epithelial transplantation for severe ocular surface diseases.
abstract: OBJECTIVE
  To determine the long-term epithelial lineage of origin of surgically removed grafts after allogeneic cultivated corneal limbal epithelial transplantation (CLET).
  
  
  METHODS
  Interventional case reports.
  
  
  METHODS
  We studied 2 eyes from 2 patients with total corneal stem cell destruction; 1 eye was from a patient with Stevens-Johnson syndrome and 1 eye had sustained chemical injury.
  
  
  METHODS
  Allogeneic cultivated corneal limbal epithelial sheets on human amniotic membrane (AM) were transplanted onto the ocular surface. Regrafting (1 eye, 42 months later) or penetrating keratoplasty (1 eye, 75 months later) were performed after the initial transplantation procedure for further visual rehabilitation.
  
  
  METHODS
  The excised grafts were subjected to clinical evaluation and to light- and transmission electron microscopy (TEM) examination and to immunohistochemical analysis.
  
  
  RESULTS
  In clinically conjunctival grafts, TEM and immunohistochemical analysis disclosed only small areas where the original cultivated corneal epithelial cells persisted. Neighboring conjunctival epithelial cells had apparently invaded a large portion of the corneal surface (keratin 3/12(-), Muc5ac(+)). In clinically corneal grafts, transplanted allogeneic cultivated corneal epithelial cells clearly survived for a long period of time (keratin 3/12(+), Muc5ac(-)); there was no infiltration by inflammatory cells, nor was there dissolution of the AM substrate.
  
  
  CONCLUSIONS
  We theorize that the process of graft opacification after allogeneic CLET is responsible for the loss of transplanted cultivated corneal epithelial cells and that this is followed by conjunctival cell invasion onto the corneal surface. The results of this study confirmed that in the clinically evaluated corneal graft, transplanted cultivated corneal epithelial cells indeed survived for a long period of time on the corneal surface and maintained ocular surface integrity, even though the transplanted cells were allogeneic.
authors.affiliation: Department of Ophthalmology, Graduate School of Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan. tnakamur@koto.kpu-m.ac.jp
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakamura
authors.forename: Chie
authors.initials: C
authors.lastname: Sotozono
authors.forename: Adams J
authors.initials: AJ
authors.lastname: Bentley
authors.forename: Sachiko
authors.initials: S
authors.lastname: Mano
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Inatomi
authors.forename: Noriko
authors.initials: N
authors.lastname: Koizumi
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Fullwood
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D002057:Burns, Chemical / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D019070:Cell Lineage; D002470:Cell Survival; D017690:Cell Transplantation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism / Q000637:transplantation / Q000648:ultrastructure; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D005126:Eye Burns / Q000139:chemically induced*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D053551:Keratin-12 / Q000378:metabolism; D053558:Keratin-3 / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology*; D008297:Male; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D055271:Mucin 5AC / Q000378:metabolism; D010641:Phenotype; D013262:Stevens-Johnson Syndrome / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D014184:Transplantation, Homologous
chemical_list: D015415:Biomarkers; D053551:Keratin-12; D053558:Keratin-3; C524678:MUC5AC protein, human; D055271:Mucin 5AC