Outcomes and DNA analysis of ex vivo expanded stem cell allograft for ocular surface reconstruction.
OBJECTIVE
To investigate the outcome of a new technique of ex vivo expanded stem cell allograft for limbal stem cell deficiency (LSCD), and to characterize the ocular surface genotype after surgery.


METHODS
Retrospective noncomparative case series.


METHODS
Ten eyes of 10 patients with profound LSCD arising from ectodermal dysplasia (3 eyes), Stevens-Johnson syndrome (3 eyes), chemical injury (2 eyes), thermal injury (1 eye), and rosacea blepharoconjunctivitis (1 eye).


METHODS
Allogeneic corneal limbal stem cells were cultured on plastic and transplanted to the recipient eye after removal of conjunctival pannus. Amniotic membrane was applied in a bandage capacity. The procedure was combined with other reconstructive surgery in 2 cases. Nine patients received systemic cyclosporin A immunosuppression, and the DNA genotype was investigated with surface impression cytology.


METHODS
Parameters of LSCD, including vascularization, conjunctivalization, inflammation, epithelial defect, photophobia, and pain.


RESULTS
The mean follow-up period was 28 months (range, 12-50). Seven of 10 eyes (70%) had improved parameters of LSCD at final follow-up and were considered successes. Four (40%) had improved visual acuity, including 3 having had further procedures for visual rehabilitation. Three patients failed to improve-1 with a thermal burn and lid deformity, 1 with Stevens-Johnson syndrome and severe dry eye, and 1 with ectodermal dysplasia who developed an epithelial defect at 26 months. DNA analysis of the first 7 cases showed no ex vivo donor stem cell DNA present beyond 9 months.


CONCLUSIONS
Ex vivo expanded stem cell allograft is a useful technique for restoring the ocular surface in profound LSCD. The absence of donor DNA beyond 9 months suggests that ongoing immunosuppression may be unnecessary and raises questions regarding the origin of the host corneal epithelium.
Daya
Sheraz M
SM
Corneo-Plastic Unit, Queen Victoria Hospital, East Grinstead, West Sussex, United Kingdom. sdaya@centreforsight.com
Watson
Adam
A
Sharpe
Justin R
JR
Giledi
Osama
O
Rowe
Andrea
A
Martin
Robin
R
James
S Elizabeth
SE
D000293:Adolescent; D000328:Adult; D000650:Amnion / Q000637:transplantation; D002478:Cells, Cultured; D002675:Child, Preschool; D003316:Corneal Diseases / Q000473:pathology / Q000601:surgery*; D004247:DNA / Q000032:analysis*; D019573:Epithelium, Corneal / Q000637:transplantation*; D005260:Female; D005838:Genotype; D006801:Humans; D007165:Immunosuppression Therapy; D016850:Limbus Corneae / Q000166:cytology*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000737:chemistry*; D014184:Transplantation, Homologous
D004247:DNA