Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation.
OBJECTIVE
It would be advantageous if cultivated human corneal endothelial cells (cHCECs) could be transplanted for the treatment of diseases caused by corneal endothelial disorders. To achieve this, a matrix that can serve as a carrier for cHCECs is needed. The present study was conducted to examine the feasibility of using amniotic membrane (AM) as a carrier for this application.


METHODS
HCECs obtained from peripheral corneal tissue were cultivated, passaged, and transplanted onto denuded AM. The cell density and morphology of the resultant cHCECs on AM were examined by light, scanning electron, and transmission electron microscopy. To determine whether these cHCEC sheets on AM carrier were functional in vivo, the cHCEC sheets on AM were transplanted onto rabbit corneas whose Descemet's membrane and endothelial cells had been completely removed. After transplantation, the corneal appearance was examined by slit lamp biomicroscopy, and corneal thickness was measured daily by pachymetry. At 7 days after surgery, the grafts were examined by light, scanning electron, and transmission electron microscopy.


RESULTS
The density of the cHCECs on AM was greater than 3000 cells/mm(2). Morphologically, the cHCEC sheets consisted of a fairly continuous layer of flat squamous polygonal endothelial cells that appeared uniform in size with tightly opposed cell junctions in vitro and in vivo after transplantation. The corneas that received transplanted cHCEC sheets had little edema and retained their thinness and transparency.


CONCLUSIONS
The cell density and morphology of cHCECs on AM were similar to those of normal corneas, and cHCECs on AM were functional in vivo. These results indicate that AM maintains HCEC morphology and function and could serve as a carrier for cHCEC transplantation.
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan. yishino@ophth.kpu-m.ac.jp
Yutaka
Y
Ishino
Yoichiro
Y
Sano
Takahiro
T
Nakamura
Che J
CJ
Connon
Helen
H
Rigby
Nigel J
NJ
Fullwood
Shigeru
S
Kinoshita
D000650:Amnion / Q000166:cytology / Q000637:transplantation*; D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques; D003316:Corneal Diseases / Q000601:surgery*; D016039:Corneal Transplantation; D003886:Descemet Membrane / Q000601:surgery*; D004728:Endothelium, Corneal / Q000166:cytology / Q000637:transplantation* / Q000648:ultrastructure; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D011817:Rabbits; D014019:Tissue Donors