Guinea pig endothelial cells in culture.

Abstract

ABSTRACTIn pursuit of a model for the studies of endothelial cells in a readily available experimental animal, culture methods and the in vitro characteristics of endothelial cells isolated from guinea pig aorta are described. Endothelial cells were harvested by trypsinization with perfusion techniques and cultured in Eagle's minimal essential medium supplemented with 10% fetal calf serum. Their passage cultivation was possible for more than six months and they have maintained the in vitro morphological characteristics of endothelial cells.The cell doubling time of the passaged endothelial cells was 115–131 hours. The growth of guinea pig aorta endothelial cells was satisfactory in Eagle's minimal essential medium containing guinea pig serum and fetal calf serum, but not horse and calf serum. Of the various concentrations of fetal calf serum in Eagle's minimal essential medium, 3% fetal calf serum did not activate cell growth and 10% fetal calf serum induced the best growth; in 20% concentration of fetal calf serum the cell growth rate decreased as compared with the rate in 10% fetal calf serum. DNA synthesis (3H‐thymidine uptake) of the cells in 20% fetal calf serum was less than that in 10% fetal calf serum. These results have no relation to cell generation nor to the cell density. With respect to the type of media supplemented with 10% fetal calf serum, Dulbecco's modified Eagle's medium, RPMI −1640 and Eagle's minimal essential medium were satisfactory for the cultivation of guinea pig aorta endothelial cells, but Medium‐199 was unsuitable. By adding endothelial cell growth supplement (75, 150 mcg) or fibroblast growth factor (100, 200 ng) to Eagle's minimal essential medium containing 3% fetal calf serum, the cell growth was stimulated approximately three times. Fibronectin‐coated wells did not activate cell growth.