November 1988
Volume 29, Issue 11
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Articles  |   November 1988
Isolation and long-term cultivation of human corneal endothelial cells.
Author Affiliations
  • K Engelmann
    Department of Cytogenetics, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Federal Republic of Germany.
  • M Böhnke
    Department of Cytogenetics, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Federal Republic of Germany.
  • P Friedl
    Department of Cytogenetics, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Federal Republic of Germany.
Investigative Ophthalmology & Visual Science November 1988, Vol.29, 1656-1662. doi:
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      K Engelmann, M Böhnke, P Friedl; Isolation and long-term cultivation of human corneal endothelial cells.. Invest. Ophthalmol. Vis. Sci. 1988;29(11):1656-1662.

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      © ARVO (1962-2015); The Authors (2016-present)

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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.

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