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Effects of insulin and insulin-like growth factor I on growth of human
leukemia cells in serum-free and protein-free medium
J Sinclair, D McClain and R Taetle
Department of Medicine, University of California, San Diego 92103.
Human myeloid leukemia cells (HL60) and malignant lymphocytes (Namalwa)
were grown in protein-free, Fe-supplemented media and used to study growth
responses to insulin and insulin-like growth factor 1 (IGF-I). HL60 cells
previously grown in serum-free medium containing microgram quantities of
insulin showed an 18-fold reduction in cumulative cell production when
grown without insulin. However, the same cells showed reduced or absent
growth stimulation with 1 to 100 ng/mL insulin or IGF- I for at least four
days following insulin deprivation, indicating that culture conditions
modified insulin and IGF-I responses. When the same cells were grown in
Fe-supplemented, protein-free medium (RPMI-Fe), insulin and IGF-I caused
dose-dependent stimulation of HL60 cell growth with half-maximal
stimulation at nanogram concentrations. Namalwa cells grown in protein-free
medium showed no response to either hormone. Radioligand binding showed the
presence of insulin and IGF-I receptors on both HL60 and Namalwa cells
grown in RPMI-Fe. HL60 cells grown in fetal bovine serum had higher, and
cells grown with microgram quantities of insulin dramatically reduced,
insulin binding. Competitive binding studies and cultures with anti-IGF-I
receptor antibody showed insulin and IGF-I stimulated growth through their
respective specific receptors. Both insulin and IGF-I stimulate growth of
some cultured human leukemia cells, but the presence of insulin or IGF-I
receptors alone does not predict growth responses. Culture conditions
affect both cellular responses and ligand binding by these hormones and
must be closely controlled to study growth responses.
Volume 72,
Issue 1,
pp. 66-72,
07/01/1988
Copyright © 1988 by The American Society of Hematology
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