title: Identifying the origin of single corneal cells by DNA fingerprinting: part I--implications for corneal limbal allografting.
abstract: OBJECTIVE
  To demonstrate that the combination of impression cytology and single cell DNA fingerprinting represents a powerful tool that is suitable for detecting transplanted cells after corneal limbal allografting.
  
  
  METHODS
  Fifty single cells were obtained by corneal impression cytology from 12 patients undergoing cataract surgery. Individual cells were isolated from samples by micromanipulation. Polymerase chain reaction and short tandem repeat profiling was used to obtain forensic standard "DNA fingerprints" from single cells. Blood samples taken at the time of impression cytology provided control "fingerprints."
  
  
  RESULTS
  Informative DNA fingerprints were obtained from all corneal samples and 66% (33 of 50 cells) of isolated single cells. Of all fingerprints obtained, most (91%, 30 of 33 fingerprints) corneal fingerprints matched corresponding blood sample fingerprints. At least one corneal fingerprint matched the corresponding blood sample fingerprint in 83% (10 of 12 patients) of the patients in the study.
  
  
  CONCLUSIONS
  This extremely specific single cell DNA fingerprinting system permits accurate identification of individual corneal epithelial cells, allowing very reliable determination of their origin, which will enable host and donor cells to be distinguished from each other after keratolimbal allografting procedures, even if the host and donor are the same sex or siblings. These DNA fingerprinting methods allow assessment of quality and quantity of donor cell survival, as well as survival time. The extreme sensitivity and accuracy of the technique means that should contamination occur, it would be identified, thus ensuring meaningful results.
authors.affiliation: Department of Ophthalmology, Leeds General Infirmary, Leeds, U.K.
authors.forename: T R
authors.initials: TR
authors.lastname: Henderson
authors.forename: I
authors.initials: I
authors.lastname: Findlay
authors.forename: P L
authors.initials: PL
authors.lastname: Matthews
authors.forename: B A
authors.initials: BA
authors.lastname: Noble
mesh_terms: D017690:Cell Transplantation; D004247:DNA / Q000032:analysis; D016172:DNA Fingerprinting / Q000379:methods*; D004847:Epithelial Cells / Q000637:transplantation*; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology; D020080:Tandem Repeat Sequences; D014019:Tissue Donors; D014184:Transplantation, Homologous
chemical_list: D004247:DNA