Nucleoside requirements for the in vitro growth of bovine aortic endothelial cells

Abstract

The nutritional needs of cultured fetal bovine aortic endothelial cells were studied with regard to their nucleotide metabolism. When Medium 199 containing calf serum was supplemented with up to 5 microgram/ml of the deoxyribo- or ribonucleosides found in DNA or RNA, the rate of endothelial cell growth increased. The effect was entirely attributable to the pyrimidine nucleosides. The combination of deoxythymidine and deoxycytidine was much more effective than either deoxyribonucleoside used alone or than the combination of uridine and cytidine. Addition of deoxythymidine and deoxycytidine (each at 1 microgram/ml) to the medium supported the growth of endothelial cell cultures from initially sparse populations (ca. 50 cells/cm2), even at low concentrations (1%) of fetal bovine serum. The pyrimidine deoxyribonucleosides on their own were unable to stimulate cell growth; other bonafide growth stimulatory factors, such as those present in serum, serum dialysates, or retinal extracts, were needed in the medium to signal the initiation of DNA synthesis and cell replication. The significance of these findings with respect to improving cell performance under in vitro conditions and controlling endothelial cell growth in vivo are discussed.

Proliferative variability of endothelial clones derived from adult bovine aorta: Influence of fibroblast growth factor and smooth muscle cell extracellular matrix

“…This indicates that the provision of an ECM coating on plastic tissue culture dishes provides a general stimulus for maintaining BAE cells in a stage of active proliferation. Because cultures that are more actively proliferating can apparently escape precocious in vitro senescence Fenselau et al, 1981;Johnson and Longenecker, 1982), it may be argued that the use of such culture conditions will extend the proliferative lifespan in culture. Although analysis of a greater number of clones will be required before any firm conclusion may be made regarding this possibility, it is apparent from the data shown in Figure 8 that those clones that proliferated at the highest rate also attained the greatest lifespans in culture.…”

“…In the following studies we have examined two mitogenic factors and have compared their influence on the clonal variability of growth of endothelial cells derived from adult bovine aorta. Previous reports have shown that the optimal proliferation of bovine aortic endothelial (BAE) cells in culture is sensitive to the type of media employed and depends in large part upon obtaining a suitable serum lot (McAuslan et al, 1979;Duthu and Smith, 1980;Fenselau et al, 1981;Gospodarowicz and Lui, 1981). Fibroblast growth factor (FGF) when used in combination with an optimum media and serum lot can increase both the growth rate at low densities and extend the proliferative lifespan (Vlodavsky et al 1979;Duthu and Smith, 1980;Johnson and Longenecker, 1982).…”

Judah Folkman, a pioneer in the study of angiogenesis

“…Tumor angiogenesis factor has since been nondestructively extracted from several tumor cell lines, and several low molecular weight angiogenic factors have been isolated, again from the Walker 256 carcinoma. These factors induced a vasoproliferative response in vivo when tested on rabbit cornea or chick CAM, and in vitro on cultured endothelial cells [ 21 – 23 ].…”

Association of methionyl-tRNA synthetase with detergent-insoluble components of the rough endoplasmic reticulum.

“…Cultures of PtK~, HeLa, mouse 3T3 fibroblast, buffalo rat liver epithelial, fetal bovine aortic endothelial (10), fetal bovine smooth muscle, and rat glial C6 cells were grown on 18 x 18 mm coverslips as described (16).…”