Sensitization of human fibroblasts to insulin-like growth factor I by serum deprivation and dexamethasone pretreatment.
Pretreatment of confluent human fibroblast cultures for two days in dexamethasone, serum free medium increased 10-20 fold the sensitivity of the cells to insulin-like growth factor I (IGF I) stimulation of amino acid uptake using the amino acid analog alpha-aminoisobutyric acid (AIB). This increased sensitivity resulted both from the use of serum free medium and the addition of dexamethasone to the serum free media. Pretreatment of the cells for 1, 2, or 3 days before assay showed that the maximum increase in sensitivity was obtained after a two day pretreatment. Pretreatment of the cells also increased their sensitivity to insulin and bovine insulin-like growth factor II stimulation of AIB uptake similar to that seen for IGF I. No consistent effect of the pretreatment was observed on either the basal level of AIB uptake or the maximal hormonal stimulation of AIB uptake. Nor was any change noted in the shape of the dose response curves. Addition of IGF I to the pretreatment medium greatly reduced the sensitivity of pretreated cells. [125I]IGF I binding studies done on suspended fibroblasts indicated that there was up to a two fold increase in the number of receptors with no increase in their affinity for IGF I. Thus, pretreatment of fibroblasts with dexamethasone and serum free medium greatly enhances their sensitivity to IGF I stimulation of AIB uptake and makes this an excellent in vitro bioassay system for IGF I.
Heath-Monnig
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Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110.
Daughaday
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D000621:Aminoisobutyric Acids / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D001769:Blood; D002460:Cell Line; D002478:Cells, Cultured; D003470:Culture Media; D003907:Dexamethasone / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D011956:Receptors, Cell Surface / Q000378:metabolism; D017451:Receptors, Somatomedin; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000166:cytology; D013002:Somatomedins / Q000494:pharmacology*
D000621:Aminoisobutyric Acids; D003470:Culture Media; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D011994:Recombinant Proteins; D013002:Somatomedins; C100049:2-aminoisobutyric acid; D007334:Insulin-Like Growth Factor I; D003907:Dexamethasone