alexa Isolation and long-term cultivation of human corneal endothelial cells.
Molecular Biology

Molecular Biology

Journal of Cell Science & Therapy

Author(s): Engelmann K, Bhnke M, Friedl P

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Abstract Human corneal endothelial cells (HCEC) were isolated by means of enzymatic treatment of excised corneas. The corneas were incubated for 1.5 hr together with a high concentration of collagenase (0.5\%), followed by a long-term incubation (up to 16 hr) using a low concentration of the enzyme (0.04\%). Endothelial cells were enriched against contaminating fibroblasts by using a selective L-valine-free medium which inhibited fibroblast growth during the first passages. Subcultures of HCEC were passaged for more than 20 generations without showing signs of senescence. Laminin and chondroitin sulfate functioned as a substrate for HCEC, promoting proliferation and allowing the cells to grow in monolayer formation. The inclusion of fibroblast growth factor (FGF) as well as chondroitin sulfate in the medium led to an additional increase in the rate of proliferation.
This article was published in Invest Ophthalmol Vis Sci and referenced in Journal of Cell Science & Therapy

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