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Updated: May 3, 2026
Growth of Human and Sheep Corneal Endothelial Cell Layers on Biomaterial Membranes
Published on: February 6, 2020
Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor
Jeffrey M A van der Krogt1, Marchien G Dallinga2, Antoon J van den Bogaerdt3
1Department of Medical Biochemistry, Amsterdam UMC, 1105 AZ Amsterdam, the Netherlands; Department of Ophthalmology, Amsterdam UMC, 1105 AZ Amsterdam, the Netherlands.
Abstract:
Studying the function of healthy corneal endothelial cells has been challenging due to a variety of culturing strategies. Here, we present a standardized protocol for culturing primary human corneal endothelial cells, isolated from research-grade corneoscleral donor tissue, without the addition of mitotic enhancers. We describe the required enzymes, medium composition, cell density, and incubation times. Additionally, we use immunofluorescence microscopy to illustrate optimal outcomes and discuss troubleshooting procedures.
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