CULTIVATION AND BEHAVIOR OF AORTIC ENDOTHELIAL-CELLS IN A MEDIUM WITH LOW SERUM CONCENTRATION

  • 1 January 1980
    • journal article
    • research article
    • Vol. 39  (11-1) , 1165-1175
Abstract
Endothelial cells isolated from calf aorta were used in the subculture no. 4-10 for experiments to establish standardized and well reproducible conditions of cultivation. The cells can be cultivated in the commercial medium Eagle-MEM [minimum essential medium] with supplements of 0.1 g L-glutamine, 5.0 g peptone, 0.5 g [human] serum albumin and 10 ml (= 1%) bovine serum/1000 ml medium (MEMPAS). Immunofluorescence showed that the cell type specific marker Factor VIII antigen was localized especially in the perinuclear region of the cells. The cells were characterized with regard to their growth behavior. The MEMPAS and the Eagle-MEM with 10% serum increases the cell number in the first 4 days of the exponential growth to the same values. The use of MEMPAS in connection with a strict cultivation regime from the deep frozen cell suspension to culture in scintillation vials guaranteed well reproducible conditions of cultivation. In 14 non-selected experiments concerning the values of the cell number on the respective day, the cultures can be divided into 2 groups which differ with statistical significance. Medium conditioned by endothelial cells (K-MEMPAS) increases the cell number and the growth rate. Endothelial cells of vessels apparently are able to produce growth factors with self-stimulating effects. At this time the endothelial cell line is stored in deep frozen state up to the 25th subculture. The endothelial cells cultivated in the described standardized conditions are useful for screening of cell type specific factors with angiogenic activity.