title: Optimization of Cultured Human Corneal Endothelial Cell Sheet Transplantation and Post-Operative Sheet Evaluation in a Rabbit Model.
abstract: To optimize cultured human corneal endothelial cell (cHCEC) sheet transplantation technique for maintenance of cHCEC viability.
  
  
  
  cHCEC sheets cultured on a collagen scaffold were covered with or without Viscoat® and exposed to humidified air in the incubator. cHCEC sheets with or without Viscoat® were transplanted into cadaveric porcine eyes by the DSAEK technique with full air tamponade and incubated for various time periods. Then cell viability was determined by using the live/dead assay kit. cHCEC sheets with Viscoat® were transplanted into rabbit eyes and the sheets were histologically evaluated before and 14 days after transplantation.
  
  
  
  A collagen scaffold and Viscoat® were effective for protecting cHCEC from damage due to air exposure in vitro. All cells died after 18 hours of air exposure in porcine eyes in Viscoat® untreated control. In contrast, Viscoat® treatment sustained full cell viability following 2 hours and could maintain approximately 80% viability after 18 hours. In a rabbit model, transplanted cHCEC sheet with Viscoat® maintained cell density at 2803 ± 229 mm(2) (18% cell loss) and expression of N-cadherin, zonula occludens-1, and actin-filament localized to cell boundary as similar as donor HCEC.
  
  
  
  Viscoat® can contribute to cHCEC protection from damage caused by exposure to air.
authors.affiliation: a Corneal Regeneration Research Team , Foundation for Biomedical Research and Innovation , Kobe , Japan.|b Department of Ophthalmology , Juntendo University School of Medicine , Tokyo , Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamaguchi
authors.affiliation: a Corneal Regeneration Research Team , Foundation for Biomedical Research and Innovation , Kobe , Japan.|c Department of Ophthalmology , University of Tokyo Graduate School of Medicine , Tokyo , Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shima
authors.affiliation: a Corneal Regeneration Research Team , Foundation for Biomedical Research and Innovation , Kobe , Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Kimoto
authors.affiliation: b Department of Ophthalmology , Juntendo University School of Medicine , Tokyo , Japan.|d Department of Ophthalmology , Juntendo University Urayasu Hospital , Chiba , Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Ebihara
authors.affiliation: b Department of Ophthalmology , Juntendo University School of Medicine , Tokyo , Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
authors.affiliation: a Corneal Regeneration Research Team , Foundation for Biomedical Research and Innovation , Kobe , Japan.|c Department of Ophthalmology , University of Tokyo Graduate School of Medicine , Tokyo , Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
mesh_terms: D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003316:Corneal Diseases / Q000473:pathology / Q000601:surgery*; D016039:Corneal Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000166:cytology / Q000637:transplantation*; D005500:Follow-Up Studies; D006801:Humans; D011184:Postoperative Period; D011817:Rabbits; D013552:Swine
chemical_list: D003094:Collagen
keywords: Air exposure; collagen; corneal endothelial cells; regenerative medicine; viscoat