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The purpose of this study were to examine the distribution of growth-promoting activity of some chelated sera and to investigate the relationship between calcium concentration in culture medium and the growth-promoting activity of the chelated sera in comparison with that of the original serum. Results showed that the chelated serum obtained by acidic resin treatment promoted colony formation, while the number of colonies decreased in cultures containing either dialyzed serum or chelated one obtained by treatment with alkaline resin. The acid-chelated serum and its lyophilized materials maintained the growth-promoting activity for 6 months or more in a deep freezer. This activity depended on the calcium concentration of culture medium in the range from 0.06 to approximately 1.5 mM. The optimal concentration of this material for growth promotion was 4 mg/ml as the protein density, which was about a quarter of that for the original serum protein. Sera from different sources treated with the acidic resin also showed similar effect on the growth. This finding strongly suggests that the acid-chelated serum and its lyophilized protein contain a keratinocyte growth factor(s).

Effects of chelated sera and alteration in calcium concentration on the growth of oral epithelial cells in primary culture.