Growth of human corneal endothelial cells in a serum-reduced medium

Cornea. 1995 Jan;14(1):62-70.


A new medium composition was established for growth of adult human corneal endothelial cells (HCEC) under serum-reduced conditions. Growth assays were performed in clonal densities with HCEC using serum-reduced culture conditions. The growth-promoting effect of different substances was tested step by step. The new serum-reduced medium was compared with the generally used medium supplemented with conventional amounts of serum. A culture medium recently described (Engelmann K, Böhnke M, Friedl P. Optimization of culture conditions for human corneal endothelial cells. In Vitro Cell Dev Biol 1989;25:1065-72) for the long-term cultivation of human corneal endothelial cells derived from adult donors was used as the starting point for the development of a new low-serum medium. It was found that the addition to the supplements ascorbic acid, insulin, selenium, transferrin, lipids, and fibroblast growth factor to the basal medium F99 (a 1:1 mixture of Ham's F12 and M199) allowed reduction of the serum content to as low as 2%. The new medium formula, called F99sr, showed an improved dose-response curve of cell growth to serum content over the range 2-40%. To ensure the maintenance of a typical endothelial cell morphology at confluence, the serum content should be 5%.

Publication types

  • Comparative Study

MeSH terms

  • Aged
  • Ascorbic Acid / pharmacology
  • Blood
  • Cell Count
  • Cell Division / drug effects
  • Cells, Cultured
  • Child
  • Culture Media / pharmacology*
  • Endothelium, Corneal / cytology*
  • Growth Substances / pharmacology
  • Humans
  • Insulin / pharmacology
  • Middle Aged


  • Culture Media
  • Growth Substances
  • Insulin
  • Ascorbic Acid