Growth of human corneal endothelial cells in a serum-reduced medium.
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%.
Engelmann
K
K
Department of Ophthalmology, University of Hamburg, Germany.
Friedl
P
P
D000368:Aged; D001205:Ascorbic Acid / Q000494:pharmacology; D001769:Blood; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D003470:Culture Media / Q000494:pharmacology*; D004728:Endothelium, Corneal / Q000166:cytology*; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D008875:Middle Aged
D003470:Culture Media; D006133:Growth Substances; D007328:Insulin; D001205:Ascorbic Acid