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Optimization of Estrogen Growth Response in MCF-7 Cells

Thomas E. Wiese, Leos G. Kral, Kathleen E. Dennis, W. Barkley Butler and S. C. Brooks
In Vitro Cellular & Developmental Biology
Vol. 28A, No. 9/10 (Sep. - Oct., 1992), pp. 595-602
Stable URL: http://www.jstor.org/stable/4296904
Page Count: 8
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Abstract

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 ($doubling time = 33 \pm 3 h$) 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 17β ($doubling time = 55 \pm 11 h$). 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 ($^{-7}10 M$). Pooled extracts of these sera were shown to contain stimulatory levels of $estradiol-17\beta$. Dextrancoated 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.

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