MCF-7 BREAST-CANCER CELLS GROWN AS MULTICELLULAR SPHEROIDS INVITRO - EFFECT OF 17-BETA-ESTRADIOL

被引:23
作者
OLEA, N
VILLALOBOS, M
DEALMODOVAR, JMR
PEDRAZA, V
机构
[1] Department of Radiology, School of Medicine, University of Granada, Granada
关键词
D O I
10.1002/ijc.2910500123
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
To obtain multicellular spheroids from MCF-7 human breast cancer cells we adhered to the following procedure: (a) limiting the adherence of cell to the substratum; (b) seeding more than the minimum number of cells; (c) guaranteeing the presence of estrogens in the culture medium. Charcoal-dextran (CD)-treated sera seemed to inhibit spheroid formation. A reduction in the concentration of CD-human sera (from 10% to 5%) added to phenol-red-free medium facilitated progress from cellular aggregates to multicellular spheroids. Once the spheroids became initiated, size increased at a rate that showed a good fit to a Gompertzian equation (A = 0.368 +/- 0.067 alpha = 0.065 +/- 0.013, r range = 0.890-0.989). Three different patterns of spheroid morphology and proliferative kinetic were defined: (a) spheroids with diameter < 200-mu-m had a constant pattern of heterogeneity in the distribution of H-3-TdR-labelled cells and in the expression of estrogen receptors; (b) spheroids 250 to 700-mu-m in diameter showed a decrease in the proportion of H-3-TdR-labelled cells accompanying inward progression (50% in the outer shell, less than 10% in a cell layer located at a depth of 150-mu-m) while, at a depth of 170-mu-m, of signs of concurrent cellular degeneration and death were apparent; and (c) spheroids with a diameter of > 750-mu-m showed a crust of viable cells uniformly labelled with thymidine without impairment of the proportion of labelled cells when progressing inward from the spheroid crust. The larger the spheroid volume, the lower its growth fraction and the longer its volume doubling time. The hormone-dependence of MCF-7 cells in forming multicellular spheroids represents a unique experimental model for assessing estrogen action on cell organization and proliferation.
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页码:112 / 117
页数:6
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