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EMBRYONIC DEVELOPMENT & STEM CELL COMPENDIUM
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All in Vitro Cells > Primary Cell card

Human corneal endothelial cells (Harvard University)

These cells are isolated from donated corneas. Confluent endothelial cultures exhibit polygonal morphology.


For endothelial cell isolation, each cornea is placed in a Petri dish containing Medium 199 and gentamicin (50 ug/mL). Under a dissecting microscope, the Descemet’s membrane, with the attached endothelium, is stripped from the stroma and placed in a 15-mL centrifuge tube containing EDTA (0.2 mg/mL) in Hanks’ balanced salt solution, pH 7.4. After incubation for 1 hour at 37°C, cells are detached by vigorous disruption with a glass pipette. Cells are then pelleted and resuspended in culture medium consisting of OPTIMEM-1, supplemented with fetal bovine serum (8%), FGF (40 ng/mL), EGF (5 ng/mL ), NGF (20 ng/mL ), ascorbic acid (20 ug/mL), human lipids (0.005%), calcium chloride (200 mg/L ), chondroitin sulfate (0.08%), RPMI-1640 multiple vitamin solution (1% ), gentamicin (50 ug/mL), and antibiotic/antimycotic solution (1:100). Medium is changed every other day until cells reach confluency (within 10 to 14 days).

See additional Stem, Progenitor & Primary Cells for: Endothelium , Eye
Primary Cell
Homo sapiens
Human corneal endothelial cells