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Ex vivo expansion and transplantation of limbal epithelial stem cells

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dc.contributor.authorShortt, A.
dc.contributor.authorSecker, G.A.
dc.contributor.authorRajan, M.
dc.contributor.authorMeligonis, G.
dc.contributor.authorDart, J.
dc.contributor.authorTuft, S.
dc.contributor.authorDaniels, J.
dc.date.issued2008
dc.description.abstractConclusions: These data demonstrate that it is possible to culture LESCs ex vivo in compliance with good manufacturing practice regulations. A set of objective outcome measures that confirm the efficiency of this technique in treating LESC deficiency is described. The widespread use of such standardized and objective outcome measures would facilitate a comparison between the different culture methods in use. Design: Prospective, noncomparative, interventional case series. Main Outcome Measures: Clinical parameters of LESC deficiency (loss of epithelial transparency, superficial corneal vascularization, epithelial irregularity, and epithelial breakdown), visual acuity, impression cytology and cytokeratin expression profiles, and in vivo confocal corneal confocal microscopy. Methods: Allogeneic (7 eyes) or autologous (3 eyes) corneal LESCs were cultured on human amniotic membrane. Tissue was transplanted to the recipient eye after superficial keratectomy. Impression cytology and confocal microscopy were performed 6 months after surgery with clinical follow-up to 13 months. Success was defined as an improvement in the defined clinical parameters of LESC deficiency, an improvement in visual acuity, the restoration of a more normal corneal phenotype on impression cytology, and the appearance of a regular hexagonal basal layer of cells on corneal confocal microscopy. Objective: To determine, using objective measures, the outcome of ex vivo cultured limbal epithelial stem cell (LESC) transplantation performed in compliance with good manufacturing practice using a novel culture system without 3T3 feeder cells. Participants: Ten eyes of 10 patients with profound LESC deficiency arising from chemical injury (4 eyes), aniridia (3 eyes), ectodermal dysplasia (1 eye), Reiger's anomaly with Pax6 haploinsufficiency (1 eye), and unknown cause (1 eye). Results: The success rate using this technique was 60% (autografts 33%, allografts 71 %). All patients with a successful outcome experienced an improvement in visual acuity of ≥ 2 lines Snellen acuity. Preoperatively, CK3+ and CK19+ cells accounted for 12± 2.4% (mean ± standard error of the mean) and 80± 2.15% of cells, respectively, whereas postoperatively these accounted for 69± 6.43% (P< 0.0001) and 30± 6.34% (P< 0.0001) of cells, respectively. Goblet cells accounted for 8 ± 1 . 19% of cells preoperatively and 1 ± 0.35% of cells postoperatively (P< 0.0001).
dc.identifier.citationOphthalmology (Rochester, Minn.), 2008; 115(11):1989-1997
dc.identifier.doi10.1016/j.ophtha.2008.04.039
dc.identifier.issn0161-6420
dc.identifier.issn1549-4713
dc.identifier.orcidSecker, G.A. [0000-0002-5109-3222]
dc.identifier.urihttps://hdl.handle.net/1959.8/154783
dc.language.isoen
dc.publisherElsevier
dc.rightsCopyright 2008 American Academy of Ophthalmology
dc.source.urihttps://doi.org/10.1016/j.ophtha.2008.04.039
dc.subjectEpithelium, Corneal
dc.subjectLimbus Corneae
dc.subjectCells, Cultured
dc.subjectStem Cells
dc.subjectHumans
dc.subjectCorneal Diseases
dc.subjectImmunosuppressive Agents
dc.subjectMicroscopy, Confocal
dc.subjectTreatment Outcome
dc.subjectCell Transplantation
dc.subjectStem Cell Transplantation
dc.subjectTransplantation, Autologous
dc.subjectTransplantation, Homologous
dc.subjectCell Count
dc.subjectProspective Studies
dc.subjectVisual Acuity
dc.subjectAdult
dc.subjectAged
dc.subjectMiddle Aged
dc.subjectFemale
dc.subjectMale
dc.titleEx vivo expansion and transplantation of limbal epithelial stem cells
dc.typeJournal article
pubs.publication-statusPublished
ror.mmsid9915911066301831

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