title: Transplantation of conjunctival epithelial cells cultivated ex vivo in patients with total limbal stem cell deficiency.
abstract: OBJECTIVE
  To report the outcomes of transplantation of autologous conjunctival epithelial cells cultivated ex vivo (EVCAU) in patients with total limbal stem cell deficiency (LSCD).
  
  
  METHODS
  EVCAU were cultivated on denuded human amniotic membrane and transplanted in 12 eyes of 10 patients with total LSCD. We evaluated the improvement in the defined clinical parameters of LSCD (loss of corneal epithelial transparency, superficial corneal neovascularization and epithelial irregularity/recurrent epithelial breakdown), vision acuity, impression cytology, immunocytochemical analysis (CK3/CK19), and the appearance of a regular hexagonal basal layer of cells on corneal confocal microscopy. Histologic and immunohistochemical features were studied in 3 corneal buttons of patients submitted to penetrating keratoplasty after EVCAU.
  
  
  RESULTS
  Cultivated conjunctival epithelium formed 4 to 5 layers with the formation of basement membrane-like structures. Immunocytochemical analysis showed positivity for CK3, CK19, MUC5AC, Ki-67, P63, and ABCG2. The improvement of the clinical parameters for this treatment in our cohort was 10 of 12 (83.3%) in a mean follow-up time of 18.5 months (range, 15-26 months), and these eyes showed an improvement in impression cytology, immunocytochemistry, and in vivo confocal analysis. Corneal buttons showed a well-formed epithelium with 5 to 6 layers, with rare cells periodic acid-Schiff+, and positivity for CK3, CK19, P63, connexin 43, and MUC5AC.
  
  
  CONCLUSIONS
  We demonstrated the preliminary results of transplantation of EVCAU for corneal surface reconstruction in cases with total LSCD. Future studies are needed to further assess the long-term efficacy of this procedure.
authors.affiliation: Ocular Surface Advanced Center, Department of Ophthalmology, Federal University of São Paulo, São Paulo, Brazil.
authors.forename: José Reinaldo S
authors.initials: JR
authors.lastname: Ricardo
authors.forename: Priscila C
authors.initials: PC
authors.lastname: Cristovam
authors.forename: Pedro A N
authors.initials: PA
authors.lastname: Filho
authors.forename: Charles C
authors.initials: CC
authors.lastname: Farias
authors.forename: Aline L
authors.initials: AL
authors.lastname: de Araujo
authors.forename: Renata R
authors.initials: RR
authors.lastname: Loureiro
authors.forename: Joyce L
authors.initials: JL
authors.lastname: Covre
authors.forename: Jeison N
authors.initials: JN
authors.lastname: de Barros
authors.forename: Telma P
authors.initials: TP
authors.lastname: Barreiro
authors.forename: Myrna S
authors.initials: MS
authors.lastname: dos Santos
authors.forename: José A P
authors.initials: JA
authors.lastname: Gomes
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000378:metabolism; D017690:Cell Transplantation; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology*; D003316:Corneal Diseases / Q000601:surgery*; D004847:Epithelial Cells / Q000637:transplantation*; D019573:Epithelium, Corneal / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D053539:Keratin-19 / Q000378:metabolism; D053558:Keratin-3 / Q000378:metabolism; D015948:Keratoplasty, Penetrating; D019394:Ki-67 Antigen / Q000378:metabolism; D016850:Limbus Corneae / Q000473:pathology; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D055271:Mucin 5AC / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D011446:Prospective Studies; D013234:Stem Cells / Q000378:metabolism / Q000473:pathology*; D014182:Transplantation, Autologous; D014792:Visual Acuity / Q000502:physiology; D055815:Young Adult
chemical_list: C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D015415:Biomarkers; D053539:Keratin-19; D053558:Keratin-3; D019394:Ki-67 Antigen; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009363:Neoplasm Proteins