title: Use of DNA polymorphisms and the polymerase chain reaction to examine the survival of a human limbal stem cell allograft.
abstract: OBJECTIVE
  The extent to which limbal epithelial stem cell allografts will repopulate the human corneal ocular surface, and the time frame over which such cells survive, are uncertain. We investigated the survival of donor-derived epithelial cells after limbal stem cell allotransplantation in a patient with bilateral limbal stem cell failure by using short tandem-repeat DNA polymorphisms to distinguish donor and recipient cells.
  
  
  METHODS
  Epithelial cells were harvested by impression cytology from the grafted eye before and at various times after transplantation. DNA was extracted and amplified by the polymerase chain reaction at an informative locus, D8S264.
  
  
  RESULTS
  Cells of donor genotype were present over the grafted areas at the time of surgery but were not detected in the central cornea until 12 weeks postoperatively, indicating that repopulation of the epithelial surface from transplanted limbal stem cells took considerable time. However, by the 20th postoperative week, only recipient-type cells were detected in the grafted eye, despite systemic immunosuppression of the recipient with azathioprine and cyclosporine.
  
  
  CONCLUSIONS
  Discrimination between donor and recipient cells on the ocular surface after limbal allotransplantation was possible using genotypic variation at DNA polymorphic sites (microsatellites). Long-term survival of donor cells after limbal transplantation did not occur in this patient. Detection of DNA polymorphisms amplified by the polymerase chain reaction is a simple, rapid, and noninvasive method of following the course of transplanted cells at the ocular surface.
authors.affiliation: Department of Opthalmology, Flinders University of South Australia, Bedford Park.
authors.forename: K A
authors.initials: KA
authors.lastname: Williams
authors.forename: H M
authors.initials: HM
authors.lastname: Brereton
authors.forename: R
authors.initials: R
authors.lastname: Aggarwal
authors.forename: P J
authors.initials: PJ
authors.lastname: Sykes
authors.forename: D R
authors.initials: DR
authors.lastname: Turner
authors.forename: G R
authors.initials: GR
authors.lastname: Russ
authors.forename: D J
authors.initials: DJ
authors.lastname: Coster
mesh_terms: D000328:Adult; D002470:Cell Survival / Q000502:physiology; D017690:Cell Transplantation / Q000502:physiology*; D003261:Contact Lenses / Q000009:adverse effects; D003316:Corneal Diseases / Q000209:etiology / Q000503:physiopathology / Q000601:surgery; D004247:DNA / Q000032:analysis*; D004848:Epithelium / Q000502:physiology / Q000637:transplantation; D005260:Female; D005838:Genotype; D006085:Graft Survival; D006801:Humans; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D016850:Limbus Corneae / Q000166:cytology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D014019:Tissue Donors; D014184:Transplantation, Homologous
chemical_list: D007166:Immunosuppressive Agents; D004247:DNA