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Rosalind M. Stewart, Stephen B. Kaye, Carl M. Sheridan, Paul S. Hiscott; A Surgical Technique for Retrieval of Whole Human Cadaveric Conjunctiva. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1926.
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To develop a surgical technique for retrieval of whole human cadaveric conjunctiva for ex vivo and in vitro research purposes.
Conjunctival tissue was obtained from human cadaveric donors within 24 hours of death. A surgical technique to excise whole conjunctival tissue was developed.Conjunctival tissue was subsequently wax embedded, sectioned and examined histologically. Conjunctival epithelial cells were also harvested with trypsin and co-cultured on an inactivated J23T3 mouse fibroblast feeder layer. Cultured cells were fixed in methanol and immunofluorescently stained for the conjunctival marker cytokeratin 19.
The eyes and surrounding tissue were first cleaned with sterile water and povidone iodine. Using aseptic technique, both eyelids were split anteriorly-posteriorly along the grey line using a number 15 scalpel and Moorfield’s forceps. A 360° limbal peritomy was performed with curved scissors and a full 360° sub-Tenon’s blunt dissection performed. The fatty tissue and Tenon’s capsule behind the conjunctiva was then divided using scissors. Eye sockets were reconstructed using cotton wool and plastic eye shields in the standard manner with good aesthetic results. Using this technique whole conjunctival tissue was successfully obtained from 14 human cadaveric donors (28 eyes). Histological analysis of the wax embedded sections displayed tarsal to limbal conjunctiva. Primary colonies of epithelial cells were cultured which stained positive for cytokeratin 19.
We have developed a technique to retrieve whole human cadaveric conjunctiva for research purposes. This enables future ex vivo and in vitro research analysis of the whole human conjunctiva.
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