Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation.
OBJECTIVE
The corneal epithelium is essential for maintaining corneal transparency, and efforts have been made to develop improved techniques for corneal epithelial transplantation in patients with total limbal failure. We evaluated the suitability of transplanted cultivated human conjunctival epithelium (HCjE) as a corneal epithelium replacement in rabbits with total corneal and limbal deficiency.


METHODS
HCjE cells, cultivated on human amniotic membrane (AM) to confluence and exposed to an air-liquid interface (air-lifted), were transplanted onto denuded rabbit corneas and monitored for 2 weeks. The cultivated HCjE sheet and the engrafted epithelium were analyzed by immunohistochemistry and transmission electron microscopy (TEM).


RESULTS
The transplanted HCjE remained transparent, smooth, and without epithelial defects during the follow-up period. Both the cultivated HCjE cells and the engrafted epithelium manifested five to six layers of stratified squamous epithelium similar in morphology to normal corneal epithelium. The basal cells expressed the putative stem cell markers (ABCG2 and P63) and hemidesmosome and desmosome component proteins. The cytokeratins (CK4, CK13, CK3, and CK12) and MUC4 were found in the engrafted epithelium. However, MUC5AC was not expressed. The results indicate that HCjE cultivated on AM has the potential to be used as an alternative corneal epithelium.


CONCLUSIONS
The transplantation of cultivated HCjE sheets is a promising technique for the treatment of eyes with limbal failure.
Tanioka
Hidetoshi
H
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan. htanioka@eye.opth.kup-m.ac.jp
Kawasaki
Satoshi
S
Yamasaki
Kenta
K
Ang
Leonard P K
LP
Koizumi
Noriko
N
Nakamura
Takahiro
T
Yokoi
Norihiko
N
Komuro
Aoi
A
Inatomi
Tsutomu
T
Kinoshita
Shigeru
S
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D017690:Cell Transplantation; D003228:Conjunctiva / Q000166:cytology*; D003316:Corneal Diseases / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D004847:Epithelial Cells / Q000637:transplantation*; D004848:Epithelium; D019573:Epithelium, Corneal / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D055264:Mucin-4; D009077:Mucins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D011817:Rabbits; D014182:Transplantation, Autologous
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; C081876:CKAP4 protein, human; C524658:MUC4 protein, human; D008565:Membrane Proteins; D055264:Mucin-4; D009077:Mucins; D009363:Neoplasm Proteins; D007633:Keratins