Fluorescent in situ hybridization (FISH) on corneal impression cytology specimens (CICS): study of epithelial cell survival after keratoplasty.
OBJECTIVE
To assess corneal epithelial cell survival after keratoplasty.


METHODS
Corneal impression cytology (CIC) was performed on sex-mismatched corneal transplants. Fluorescent in situ hybridization (FISH) with sex chromosome-specific probes was performed to identify epithelial cell mosaicism and therefore allocate the donor or recipient origin of the cells. Twenty-four samples of corneal epithelial cells derived from 21 transplanted patients were analyzed. All patients received post-operative treatment using dexamethasone eye drops, with progressive tapering over 18 months, and nine patients also received 2% cyclosporine eye drops.


RESULTS
Out of the 24 samples reaching quality criteria, sex mosaicism was found in 13, demonstrating the presence of donor-derived cells at the center of the graft for at least 211 days post keratoplasty. Kaplan-Meier analysis established a median survival of donor corneal epithelial cells of 385 days. Although not statistically significant, the disappearance of donor cells seemed to be delayed and the average number of persistent cells appeared to be greater when 2% cyclosporine was used topically as an additional immunosuppressive therapy.


CONCLUSIONS
The combination of corneal impressions and FISH analysis is a valuable tool with negligible side effects to investigate the presence of epithelial cell mosaicism in sex-mismatched donor transplants. Epithelial cells survived at the center of the graft with a median survival of more than one year, suggesting slower epithelial turnover than previously described.
Department of Ophthalmology, Fondation Rothschild and Hôpital Bichat AP-HP, Paris, France.
Muriel
M
Catanese
Cornel
C
Popovici
Hélène
H
Proust
Louis
L
Hoffart
Frédéric
F
Matonti
Isabelle
I
Cochereau
John
J
Conrath
Eric E
EE
Gabison
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002470:Cell Survival; D002648:Child; D041321:Chromosomes, Human, X / Q000378:metabolism; D041322:Chromosomes, Human, Y / Q000378:metabolism; D003316:Corneal Diseases / Q000601:surgery*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D005260:Female; D006085:Graft Survival / Q000502:physiology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015948:Keratoplasty, Penetrating; D008297:Male; D008875:Middle Aged; D009030:Mosaicism; D014019:Tissue Donors