Long-term phenotypic study after allogeneic cultivated corneal limbal epithelial transplantation for severe ocular surface diseases.
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.
Nakamura
Takahiro
T
Department of Ophthalmology, Graduate School of Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan. tnakamur@koto.kpu-m.ac.jp
Sotozono
Chie
C
Bentley
Adams J
AJ
Mano
Sachiko
S
Inatomi
Tsutomu
T
Koizumi
Noriko
N
Fullwood
Nigel J
NJ
Kinoshita
Shigeru
S
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
D015415:Biomarkers; D053551:Keratin-12; D053558:Keratin-3; C524678:MUC5AC protein, human; D055271:Mucin 5AC