An optimized culture medium for human vascular endothelial cells from umbilical cord veins.

Various polypeptide growth factors, culture substrates, basal media, sera and further supplements were assayed for improvement of growth of human vascular endothelial cells from umbilical cord veins. The resulting optimized medium consisted of gelatinized culture substrates, a mixture (1:1) of Iscove's MDM and Ham's F12 basal media supplemented with 20% newborn calf serum, 500 ng/ml crude fibroblast growth factor, 20 ng/ml epidermal growth factor, 5 μg/ml transferrin, 5 μg/ml insulin and 10 μg/ml heparin. The medium allowed long term cultivation of HUVEC up to 45 generations with maximal cell densities of about 10(5) cells per cm(2) and a minimal doubling time of about 14 hours at low cell densities.