title: [Comparison between amniotic membrane with and without epithelium as a support for human limbal epithelial cells cultured ex vivo].
abstract: OBJECTIVE
  To evaluate the efficacy and ultrastructural aspects of human limbal epithelial cells cultured on amniotic membrane (AM) with and without epithelium.
  
  
  METHODS
  Limbal epithelial cell cultures were established from cadaveric cor neo-scleral rim explants derived from 6 different donors. The explants from each donor were placed under 3 different groups: on human preserved AM with epithelium (Group 1), AM deepithelialized with trypsin (Group 2) and control (Group 3). The epithelial cell migration was evaluated under phase contrast microscopy. After 15 days, the amniotic membrane with cells cultures were removed and submitted to scanning and transmission electron microscopy to check for epithelial migration and adhesion.
  
  
  RESULTS
  All epithelial cell cultures from the controls grew over the botton of the culture plate wells until reaching confluence. Epithelial cultures grew over all but one denuded amniotic membrane. In the group amniotic membrane with epithelium, epithelial cell growing was observed only in 1 well.
  
  
  CONCLUSIONS
  Using this model, denuded amniotic membrane appeared to be the best substrate for epithelial cell migration and adhesion comparing to amniotic membrane with epithelium. Removal of amniotic membrane epithelial seems to be an important step for establishing limbal epithelial cell culture on amniotic membrane.
authors.affiliation: Centro Avançado de Superfície Ocular, Instituto da Visão, Departamento de Oftalmologia, Universidade Federal de São Paulo, SP, Brasil.
authors.forename: Joyce Luciana
authors.initials: JL
authors.lastname: Covre
authors.forename: Renata Ruoco
authors.initials: RR
authors.lastname: Loureiro
authors.forename: Priscila Cardoso
authors.initials: PC
authors.lastname: Cristovam
authors.forename: José Reinaldo Da Silva
authors.initials: JR
authors.lastname: Ricardo
authors.forename: Edna Freymuller
authors.initials: EF
authors.lastname: Haapalainen
authors.forename: José Álvaro Pereira
authors.initials: JÁ
authors.lastname: Gomes
mesh_terms: D000650:Amnion; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission