title: Transplantation of human embryonic stem cells onto a partially wounded human cornea in vitro.
abstract: OBJECTIVE
  The aim of this study was to investigate whether cells originating from human embryonic stem cells (hESCs) could be successfully transplanted onto a partially wounded human cornea. A second aim was to study the ability of the transplanted cells to differentiate into corneal epithelial-like cells.
  
  
  METHODS
  Spontaneously, differentiated hESCs were transplanted onto a human corneal button (without limbus) with the epithelial layer partially removed. The cells were cultured on Bowman's membrane for up to 9 days, and the culture dynamics documented in a time-lapse system. As the transplanted cells originated from a genetically engineered hESC line, they all expressed green fluorescent protein, which facilitated their identification during the culture experiments, tissue preparation and analysis. To detect any differentiation into human corneal epithelial-like cells, we analysed the transplanted cells by immunohistochemistry using antibodies specific for CK3, CK15 and PAX6.
  
  
  RESULTS
  The transplanted cells established and expanded on Bowman's membrane, forming a 1-4 cell layer surrounded by host corneal epithelial cells. Expression of the corneal marker PAX6 appeared 3 days after transplantation, and after 6 days, the cells were expressing both PAX6 and CK3.
  
  
  CONCLUSIONS
  This shows that it is possible to transplant cells originating from hESCs onto Bowman's membrane with the epithelial layer partially removed and to get these cells to establish, grow and differentiate into corneal epithelial-like cells in vitro.
authors.affiliation: Unit of Clinical Sciences, Department of Obstetrics and Gynecology, Sahlgrenska University Hospital, University of Gothenburg, Göteborg, Sweden. charles.hanson@obgyn.gu.se
authors.forename: Charles
authors.initials: C
authors.lastname: Hanson
authors.forename: Thorir
authors.initials: T
authors.lastname: Hardarson
authors.forename: Catharina
authors.initials: C
authors.lastname: Ellerström
authors.forename: Markus
authors.initials: M
authors.lastname: Nordberg
authors.forename: Gunilla
authors.initials: G
authors.lastname: Caisander
authors.forename: Mahendra
authors.initials: M
authors.lastname: Rao
authors.forename: Johan
authors.initials: J
authors.lastname: Hyllner
authors.forename: Ulf
authors.initials: U
authors.lastname: Stenevi
mesh_terms: D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D065306:Corneal Injuries; D053595:Embryonic Stem Cells / Q000166:cytology / Q000637:transplantation*; D019573:Epithelium, Corneal / Q000166:cytology / Q000378:metabolism; D005131:Eye Injuries / Q000378:metabolism / Q000628:therapy*; D005136:Eye Proteins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D053569:Keratin-15 / Q000378:metabolism; D053558:Keratin-3 / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D033581:Stem Cell Transplantation; D016610:Tissue Embedding; D016707:Tissue Fixation; D014949:Wounds, Nonpenetrating / Q000378:metabolism / Q000628:therapy*
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D053569:Keratin-15; D053558:Keratin-3; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D049452:Green Fluorescent Proteins