Title
Standardized limbal epithelial stem cell graft generation and transplantation Standardized limbal epithelial stem cell graft generation and transplantation
Author
Faculty/Department
Faculty of Medicine and Health Sciences
Publication type
article
Publication
New York, N.Y. ,
Subject
Human medicine
Source (journal)
Tissue engineering: part C: methods. - New York, N.Y.
Volume/pages
16(2010) :5 , p. 921-927
ISSN
1937-3384
1937-3392
ISI
000282223800012
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Purpose: To describe a standardized, xenogenic-free protocol for the manufacture of limbal epithelial stem cell grafts and a no touch surgical technique for its standardized transplantation. Setting: Antwerp University Hospital, Antwerp, Belgium. Methods: The limbo-amnion composite graft is generated by cultivating limbal epithelial stem cells on a standardized (thermolysin treated and spongy layer removed) amniotic membrane, stretched within an interlockable amnion ring. The cells are cultured in CnT-20 medium with the addition of 1% human AB serum for a period of 2 weeks. Fibrin glue is applied to the surgically prepared recipient's cornea and in one fluid motion, the composite graft within the amnion ring construct is transferred from culture and positioned onto the graft bed. The required size is cut out at the level of the limbus by means of a trephine and/or microsurgical scissors. Results: The lightweight, plastic interlockable ring offered stability to the graft during culture, transport, and transplantation. The use of the standardized amniotic membrane, within the amnion ring construct, improves reproducibility of the results and therefore heralds elective surgery. Rapid transplantation of a wrinkle-free graft, using a sutureless, no touch technique was achieved and this allowed precise tailoring of the graft to the recipient bed. Conclusion: This is the first time a standardized, clinical grade protocol has been described for manufacturing limbal epithelial grafts with an efficient surgical technique that prevents postsurgical graft shrinkage and improves corneal integration. The quick, sutureless, and manipulation-free technique ensured transplantation of viable, proliferating limbal epithelial stem cells.
E-info
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