Experimental transplantation of cultured human limbal and amniotic epithelial cells onto the corneal surface.
OBJECTIVE
Tissue-cultured corneal epithelial transplantation is a novel procedure that uses tissue-cultured epithelial cells to restore severely damaged ocular surfaces. In this study, we used tissue-cultured human limbal and amniotic epithelial cells as donor cells to investigate the feasibility of this procedure for reestablishment of a damaged ocular surface in experimental conditions.


METHODS
Primary human limbal epithelial cultures were established from banked limbal tissue. Amniotic epithelial cells were isolated from serologically screened human placenta and maintained in a specialized nutrient medium. Suspended cells (5 x 10(5)/ml) were seeded onto the concave surface of collagen corneal shields and incubated at 37 degrees C for 2-3 days. These cell-covered shields were then placed on a denuded stromal surface in organ culture and on New Zealand albino rabbit ocular surfaces that had the native epithelium previously removed. Specimens were collected 24, 48, 72, and 96 h later from organ-cultured corneal buttons and recipient animals, processed, and evaluated histologically.


RESULTS
The cells grown on the collagen shield were spread uniformly and unpolarized after 48 h in culture. They were repolarized and tightly adhered to the recipient corneal stroma 24 h after transplantation, as demonstrated by formation of cell-substrate hemidesmosomes (HDs) and donor-specific antigen immunostaining. The donor cells were retained in six of 15 rabbits receiving limbal cells and four of 12 rabbits receiving amniotic cells for as long as 10 days after surgery.


CONCLUSIONS
Cultured human limbal and amniotic epithelial cells can be successfully transplanted onto a denuded corneal surface where they adhere tightly to underlying stroma by hemidesmosomes.
He
Y G
YG
Department of Ophthalmology, The University of Texas Southwestern Medical Center at Dallas, 75235-9057, USA. yhe@mednet.swmed.edu
Alizadeh
H
H
Kinoshita
K
K
McCulley
J P
JP
D000650:Amnion / Q000166:cytology*; D000818:Animals; D001324:Autoantigens / Q000032:analysis; D002352:Carrier Proteins; D002448:Cell Adhesion; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen; D003316:Corneal Diseases / Q000473:pathology / Q000601:surgery*; D003319:Corneal Stroma / Q000601:surgery* / Q000648:ultrastructure; D017077:Culture Media, Conditioned; D003598:Cytoskeletal Proteins; D003896:Desmosomes / Q000648:ultrastructure; D000071821:Dystonin; D004847:Epithelial Cells / Q000737:chemistry / Q000637:transplantation*; D019084:Fluorescent Antibody Technique, Indirect; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D009924:Organ Culture Techniques; D011817:Rabbits; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D017077:Culture Media, Conditioned; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D015395:Histocompatibility Antigens Class I; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097805:Collagen Type XVII; C486268:DST protein, human