title: Coculture of autologous limbal and conjunctival epithelial cells to treat severe ocular surface disorders: long-term survival analysis.
abstract: BACKGROUND
  Cultivated limbal epithelium for reconstruction of corneal surface is a well-established procedure; however, it is not adequate for damage which also extensively involves the conjunctiva. In severe cases of ocular surface damage that warrant additional conjunctival transplantation apart from cultivated limbal stem cell transplantation, we describe the long-term survival of a novel method of cocultivating autologous limbal and conjunctival epithelium on a single substrate.
  
  
  METHODS
  Forty eyes of 39 patients with severe limbal stem cell deficiency and conjunctival scarring or symblepharon underwent transplantation of autologous cocultivated epithelium on human amniotic membrane. A ring barrier was used to segregate the central limbal and peripheral conjunctival epithelia in vitro. Patients were followed up at regular intervals to assess stability of the ocular surface, defined by absence of conjunctivalization into the central 4 mm of the cornea and absence of diffuse fluorescein staining. Penetrating keratoplasty (PKP) was subsequently performed, where indicated, in patients with surface stability.
  
  
  RESULTS
  The cumulative survival probability was 60% at 1 year and 45% at 4 years by Kaplan-Meier analysis (mean follow-up duration: 33 ± 29 months, range: 1-87 months). Best-corrected visual acuity improved to greater than 20/200 in 38% eyes at the last follow-up, compared with 5% eyes before surgery. Immunohistochemistry in five of the corneal buttons excised for PKP showed an epithelial phenotype similar to cornea in all five.
  
  
  CONCLUSIONS
  Synchronous use of cultured limbal and conjunctival epithelium offers a feasible alternative and a simpler one-step surgical approach to treat severe ocular surface disorders involving limbus and conjunctiva.
authors.affiliation: Cornea and Anterior Segment Services; Sudhakar and Sreekanth Ravi Stem Cell Biology Laboratory, C-TRACER, Hyderabad Eye Research Foundation, L V Prasad Eye Institute, Hyderabad, India.
authors.forename: Sandhya V
authors.initials: SV
authors.lastname: Subramaniam
authors.forename: Kunjal
authors.initials: K
authors.lastname: Sejpal
authors.forename: Anees
authors.initials: A
authors.lastname: Fatima
authors.forename: Subhash
authors.initials: S
authors.lastname: Gaddipati
authors.forename: Geeta K
authors.initials: GK
authors.lastname: Vemuganti
authors.forename: Virender S
authors.initials: VS
authors.lastname: Sangwan
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002648:Child; D018920:Coculture Techniques; D003228:Conjunctiva / Q000166:cytology / Q000637:transplantation*; D003316:Corneal Diseases / Q000601:surgery*; D016039:Corneal Transplantation / Q000379:methods*; D019573:Epithelium, Corneal / Q000166:cytology*; D005260:Female; D005500:Follow-Up Studies; D006085:Graft Survival; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D008297:Male; D008875:Middle Aged; D012189:Retrospective Studies; D013997:Time Factors; D014182:Transplantation, Autologous; D016896:Treatment Outcome; D014792:Visual Acuity; D055815:Young Adult
references.citation: Cornea. 2003 Jul;22(5):478-81
references.pmid: 12827056
references.citation: Br J Ophthalmol. 2011 Nov;95(11):1525-9
references.pmid: 21890785
references.citation: Curr Opin Ophthalmol. 2005 Aug;16(4):233-40
references.pmid: 16000896
references.citation: Transplantation. 2004 Jun 15;77(11):1729-34
references.pmid: 15201674
references.citation: J Postgrad Med. 2006 Oct-Dec;52(4):257-61
references.pmid: 17102542
references.citation: Am J Ophthalmol. 2008 Aug;146(2):266-275
references.pmid: 18514608
references.citation: Invest Ophthalmol Vis Sci. 2010 Feb;51(2):758-64
references.pmid: 19643956
references.citation: Nat Protoc. 2010 Aug;5(8):1470-9
references.pmid: 20671730
references.citation: Exp Eye Res. 2002 Apr;74(4):537-45
references.pmid: 12076097
references.citation: Am J Ophthalmol. 2005 Aug;140(2):223-30
references.pmid: 16023069
references.citation: Lancet. 1997 Apr 5;349(9057):990-3
references.pmid: 9100626
references.citation: Arch Ophthalmol. 2007 Oct;125(10):1337-44
references.pmid: 17923540
references.citation: Arch Ophthalmol. 2005 Mar;123(3):334-40
references.pmid: 15767475
references.citation: Clin Ophthalmol. 2008 Sep;2(3):489-502
references.pmid: 19668747
references.citation: Biosci Rep. 2001 Aug;21(4):481-9
references.pmid: 11900323
references.citation: Ophthalmic Surg Lasers Imaging. 2006 Sep-Oct;37(5):378-82
references.pmid: 17017197
references.citation: Ophthalmology. 2004 Jan;111(1):38-44
references.pmid: 14711712
references.citation: Indian J Ophthalmol. 2006 Mar;54(1):29-34
references.pmid: 16531667
references.citation: Ophthalmology. 1982 Sep;89(9):1072-81
references.pmid: 7177571
references.citation: Cell Tissue Bank. 2004;5(3):161-75
references.pmid: 15509905
references.citation: Ophthalmology. 2009 Jun;116(6):1048-56
references.pmid: 19394701
references.citation: Cornea. 1999 Mar;18(2):164-71
references.pmid: 10090361
references.citation: Mol Vis. 2007 Sep 18;13:1711-21
references.pmid: 17960109
references.citation: N Engl J Med. 2010 Jul 8;363(2):147-55
references.pmid: 20573916