The successful culture and autologous transplantation of rabbit oral mucosal epithelial cells on amniotic membrane.
OBJECTIVE
To determine the feasibility of using human amniotic membrane (AM) as a substrate for culturing oral epithelial cells and to investigate the possibility of using autologous cultivated oral epithelial cells in ocular surface reconstruction.


METHODS
An ocular surface injury was created in one eye of each of eight adult albino rabbits by a lamellar keratectomy, and a conjunctival excision was performed, including and extending 5 mm outside the limbus. Oral mucosal biopsy specimens were obtained from these eight adult albino rabbits and cultivated for 3 weeks on a denuded AM carrier. The cultivated epithelium was examined by electron microscopy (EM) and immunohistochemically labeled for several keratins. At 3 to 4 weeks after the ocular surface injury, the conjunctivalized corneal surfaces of the eight rabbits were surgically reconstructed by transplanting the autologous cultivated oral epithelial cells on the AM carrier.


RESULTS
The cultivated oral epithelial sheet had four to five layers of stratified, well-differentiated cells. EM revealed that the epithelial cells were very similar in appearance to those of normal corneal epithelium, had numerous desmosomal junctions, and were attached to a basement membrane with hemidesmosomes. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin-3 in the cultivated oral epithelial cells. Corneas that were grafted with the cultivated oral epithelial cells on an AM carrier were clear and were all epithelialized 10 days after surgery.


CONCLUSIONS
Cultures of oral epithelial cells can be generated to confluence on AM expanded ex vivo from biopsy-derived oral mucosal tissue. Autologous transplantation was performed with these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. Autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction. The long-term outcome of such transplantation is not yet clear, and its feasibility in clinical use should be evaluated further.
Nakamura
Takahiro
T
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto 602-0841, Japan. tnakamur@ophth.kpu-m.ac.jp
Endo
Ken-Ichi
K
Cooper
Leanne J
LJ
Fullwood
Nigel J
NJ
Tanifuji
Noriko
N
Tsuzuki
Masakatsu
M
Koizumi
Noriko
N
Inatomi
Tsutomu
T
Sano
Yoichiro
Y
Kinoshita
Shigeru
S
D000650:Amnion; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D065306:Corneal Injuries; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D005131:Eye Injuries / Q000601:surgery*; D005240:Feasibility Studies; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007633:Keratins / Q000378:metabolism; D009061:Mouth Mucosa / Q000166:cytology*; D011817:Rabbits; D014182:Transplantation, Autologous
D007633:Keratins