The Quantitative Utilization of Amino Acids and Glucose and Contact Inhibition of Growth in Cultures of the Human Diploid Cell, WI-38

Abstract

There are many reports in the literature showing that contact inhibition of growth is affected by the culture medium. A quantitative study of amino acid and glucose uptake by the human diploid cell line, WI-38 was carried out to determine more precisely what effect nutritional factors have on contact inhibition of growth. Eagle's minimal essential medium (MEM) was found to support higher cell yields than Eagle's basal medium (BME) and for growth to continue beyond 96 h a medium change was essential. However, analysis of the used growth media showed that neither amino acids nor glucose were fully depleted after 96 h. The rate of glucose utilization was in the range 65-100µg/mg dry wt./h and this agreed very closely with the results of other authors. The pattern of amino acid uptake also closely resembled that for other cell lines except that the utilization of cystine was higher. The nutritional requirements were further studied as the results from the medium analyses failed to explain the growth-promoting activity of MEM. Daily medium changes greatly increased cell yields even though the medium nutrients were not exhausted. This effect was dependent upon fresh medium being used and the only medium component found to be of importance was the amino acid complement. These results are discussed in relation to the low saturation density of diploid cells in culture and a possible explanation is proposed in terms of differences in the cell membrane between normal and altered cells.