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Maximilian Schultheiss, Berenike Kunzmann, Christian Klameth, Karl-Ulrich Bartz-Schmidt, Martin Stephan Spitzer; Establishment Of A Porcine Corneal Endothelial Organ Culture Model. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1464. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Human corneas are used for transplantation and are not available for research. Due to dedifferentiation investigations with cultured endothelial cells can be misleading. An organotypic culture of pig corneas, which show comparable endothelial cell death rates like cultured human corneas therefore would be very desirable.
Fresh pig eyes were prepared under sterile conditions as “corneoscleral buttons”, “corneal buttons” and “split corneal buttons” (new preparation method) and cultivated for 15 days. Morphology of the endothelial cell layer was observed by light microscopy on day 1, 8 and 15. On day 15 staining with trypan blue and alizarin red S was performed. Photographs were taken and evaluated in a randomized, blinded manner. The morphology of the corneal endothelium and the number of endothelial cells per mm2 was analyzed.
After 15 days of cultivation the endothelial cell layer was only maintained in corneal buttons and split corneal buttons. Alizarin red S stained dots and the existence of polymorphisms like pleomorphism, rosette figures and reformation figures were less frequent in split corneal buttons than in corneal buttons. Loss of endothelial cells was significantly greater in corneal buttons (663± 189 cells/mm2 respectively15.9%) than in split corneal buttons (443± 190 cells/mm2 respectively 11.1%).
The new preparation method of split corneal buttons allows the cultivation of porcine corneas for two weeks with comparable cell death rates like in cultured human tissue. This is therefore a simple and highly reliable method model to investigate corneal endothelial cells.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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