Estrogen mitogenic action. III. Is phenol red a 'red herring'?

Jorge E. Moreno-Cuevas, David A. Sirbasku*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)


The reported estrogenic action of phenol red and/or its lipophilic contaminants has led to the widespread use of indicator-free culture medium to conduct endocrine studies in vitro. Because we have recently developed methods to measure large-magnitude estrogen effects in the tissue culture medium containing phenol red, we concluded that the indicator issue required further evaluation. To do this, we selected nine estrogen receptor positive (ER+) cell lines representing four target tissues and three species. We investigated phenol red using five different experimental protocols. First, 17β-estradiol (E2) responsive growth of all nine ER+ cells lines was compared in the medium with and without the indicator. Second, using representative lines we asked if phenol red was mitogenic in the indicator-free medium. The dose-response effects of phenol red were compared directly to those of E2. Third, we asked if tamoxifen-inhibited growth equally in phenol red-containing and indicator-free medium. This study was based on a report indicating that antiestrogen effects should be seen only in phenol red-containing medium. Fourth, we asked if phenol red displaced the binding of 3H-E2 using ER+ intact human breast cancer cells. Fifth, we compared E2 and phenol red as inducers of the progesterone receptor using a human breast cancer cell line. All the experiments presented in this report support the conclusion that the concentration of phenol red contaminants in a standard culture medium available today is not sufficient to cause estrogenic effects. In brief, our studies indicate that the real issue of how to demonstrate estrogenic effects in culture resides elsewhere than phenol red. We have found that the demonstration of sex steroid hormone-mitogenic effects in culture depends upon conditions that maximize the effects of a serum-borne inhibitor(s). When the effects of the inhibitor are optimized, the presence or absence of phenol red makes no everyday difference to the demonstration of estrogen mitogenic effects with several target cell types from diverse species.

Original languageEnglish
Pages (from-to)447-464
Number of pages18
JournalIn Vitro Cellular and Developmental Biology - Animal
Issue number7
Publication statusPublished - 1 Jan 2000
Externally publishedYes

Bibliographical note

Funding Information:
We gratefully acknowledge the expert technical assistance of Mrs. Juanita Jenkins. This research was supported by grants DAMD17-94-J-4473, DAMD17-98-1-8337, and DAMD17-99-1-9405 from the Department of Defense, US Army Medical Research and Materiel Command, Breast Cancer Research Program, grants-in-aid from the Women's Fund for Health Education Research, Houston, Texas, a grant-in-aid from the Houston Texas Chapter of the Susan G. Komen Breast Cancer Foundation, and a grant-in-aid from the Cancer Federation, Banning, California

Copyright 2017 Elsevier B.V., All rights reserved.

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology


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