Optimization of estrogen growth response in MCF-7 cells.

Wiese TE, Kral LG, Dennis KE, Butler WB, Brooks SC
In Vitro Cell Dev Biol. 1992 28A (9-10): 595-602

PMID: 1429362 · DOI:10.1007/BF02631033

The factors involved in estradiol-17 beta induced growth stimulation of MCF-7 human breast cancer cells have been examined. Wild type MCF-7 cells (and clone E3) were shown to undergo slow growth in phenol-red-free medium containing specific calf sera. The E3 clone was used to document a mean 6-day growth stimulation of 3.35-fold (doubling time = 33 +/- 3 h) in cultures supplemented with 10(-11) M estradiol-17 beta. The serum batch utilized in the culture medium is most important in acquiring significant growth stimulation of MCF-7 cells by estradiol-17 beta. Regardless of the absence of phenol-red, only selected sera (2 out of 14 tested) supported minimal growth of MCF-7 cells in the absence of added estradiol 17 beta (doubling time = 55 +/- 11 h). When a calf-serum-supplemented culture failed to display a complete growth response to estradiol-17 beta, it was due to the rapid growth of the cells in the control (minus estradiol-17 beta) flasks. Sera that promoted shorter doubling times for MCF-7 cells cultured in the absence of estradiol-17 beta were rendered less supportive of growth if treated with dextran-coated charcoal or when cultures were supplemented with the estrogen antagonist ICI 164,384 (10(-7) M). Pooled extracts of these sera were shown to contain stimulatory levels of estradiol-17 beta. Dextran-coated charcoal treatment of sera removed or deactivated factors (other than estradiol-17 beta) which were not only required for the growth of MCF-7 cells, but were necessary for estrogen-stimulated growth. Varying the serum-containing medium, buffer, and nutrient mix or the addition of insulin has no effect on the growth response of these cells to estradiol-17 beta. These investigations document the culture conditions required to produce a maximal and consistent proliferative effect of E2 on MCF-7 cells without exposing the serum constituent to damaging chemical or absorbent agents.

MeSH Terms (10)

Blood Breast Neoplasms Cell Division Culture Media Estradiol Estrogen Antagonists Humans Insulin Polyunsaturated Alkamides Tumor Cells, Cultured

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