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Jie Zhang, Amatul Mateen Ahmad, Charles N McGhee, Dipika V. Patel; Characterisation of corneal transition zone explant cultures. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1374. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal endothelial cells have extremely limited ability to renew themselves; excessive loss through trauma or disease may result in blindness. Current treatment is only by corneal transplantation. Unfortunately, human donor cornea supply is limited and alternatives are required for the timely treatment of common corneal endothelial diseases. Adult stem cells have recently been identified in the ‘Transition zone’ in the periphery of the corneal endothelium. We aim to determine the potential of these cells for corneal endothelial transplants by characterising the ability of the Transition zone to self-renew and regenerate endothelial cells in culture.
An in vitro explant culture protocol for the human Transition Zone was established. The percentage of actively dividing cells over a 5 day period was assessed using EdU incorporation into newly synthesized DNA. The expression of stem cell and differentiated endothelial markers was assessed using immunohistochemistry and confocal microscopy.
Cells from each 0.4 cm2 piece of transition zone explant took an average of one month to reach confluency in a well of a 12 well plate (3.8 cm2). Subsequently, these cells were passaged up to 11 times before senescence. An average of 57% of cells incorporated EdU over a 5 day period at 90% confluency. The transition zone cells expressed the stem cell markers Nestin, Oct3/4, and the corneal endothelial markers ZO-1 and Na/K ATPase.
Corneal transition zone cells can proliferate and differentiate into corneal endothelial cells in vitro, demonstrating their potential to be developed into stem cell therapies for corneal endothelial diseases.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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