title: Transplantation of a sheet of human corneal endothelial cell in a rabbit model.
abstract: OBJECTIVE
  To develop a novel method for constructing a sheet of human corneal endothelial cells (HCECs) and examine the properties of the HCEC sheet.
  
  
  METHODS
  HCECs were cultured on a cell culture insert for a week; ethylenediamine tetraacetic acid was applied from the bottom of the cell culture insert to attenuate the attachment of HCECs. The sheet of HCECs was constructed by bluntly detaching the cell sheet with a spatula. HCEC cell sheets were placed on the posterior surface of excised rabbit corneal buttons and transplanted onto the corneal beds of donor rabbits. In two eyes from the HCEC sheet group, cultured HCECs were labeled with PKH26 to observe the localization of HCECs after transplantation.
  
  
  RESULTS
  Cultured HCECs could be bluntly detached en bloc from the bottom of a culture insert. Immunostaining for ZO-1, Na+, K+-ATPase, laminin, fibronectin, and type IV collagen was positive in the cell sheet. The average cell density in a HCEC sheet was 2,425 cells/mm(2). After HCEC sheet transplantation, corneal edema decreased much earlier in the HCEC group than in the control group. In the HCEC sheet group, the monolayer of continuous cells attached to the posterior surface of the transplanted rabbit cornea and the posterior surface of transplanted cornea was covered with PKH26-labeled cells. The average endothelial cell density in the HCEC sheet group seven days postoperatively was 2,244 cells/mm(2).
  
  
  CONCLUSIONS
  This technique for producing an HCEC sheet might be useful in regenerative medicine for the cornea and reconstruction of the corneal endothelium.
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
authors.forename: Kouichiro
authors.initials: K
authors.lastname: Hitani
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yokoo
authors.forename: Norihiko
authors.initials: N
authors.lastname: Honda
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Usui
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
authors.forename: Shiro
authors.initials: S
authors.lastname: Amano
mesh_terms: D000818:Animals; D000869:Anterior Eye Segment / Q000473:pathology; D002452:Cell Count; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D015715:Corneal Edema / Q000473:pathology; D003584:Cytological Techniques; D004492:Edetic Acid / Q000494:pharmacology; D004728:Endothelium, Corneal / Q000166:cytology* / Q000473:pathology / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D011817:Rabbits; D014183:Transplantation, Heterologous
chemical_list: D004492:Edetic Acid