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Cell Biology International (2001) 25, 1021–1024 (Printed in Great Britain)

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EFFECT OF PHENOL RED AND STEROID HORMONES ON CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR IN MOUSE ENDOMETRIAL EPITHELIAL CELLS

L.L. Tsang^a, L.N. Chan^a, C.Q. Liu^b and H.C. Chan^a,f1

^aEpithelial Cell Biology Research Center, Department of Physiology, The Chinese University of Hong Kong
^bShanghai Institute of Planned Parenthood Research, Shanghai, China

Abstract

Previous studies have demonstrated that cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-mediated Cl−channel found in most epithelia including reproductive tract, could be regulated by various culture conditions. The present study further investigated the effect of phenol red, a pH indicator widely used in growth medium, and steroid hormones, present in the supplement fetal bovine serum (FBS), on primary cultured endometrial epithelial cells by monitoring ion channel activities using the short-circuit current technique. When compared to the results obtained with normal medium supplemented with regular FBS, the forskolin-stimulated ISC, presumably mediated by CFTR, obtained in phenol red-free medium was significantly reduced, from 16.95±1.53μA/cm2(control) to 9.72±0.89μA/cm2(medium without phenol red, P< 0.05). The forskolin-activated ISCwas further attenuated to 5.29±0.46μA/cm2in the phenol red-free medium when supplemented with charcoal/ dextran-treated FBS where steroid hormones were removed. Our data suggest that phenol red and steroid hormones present in culture medium and FBS supplement, respectively, may somehow upregulate CFTR expression in vitro. Our study demonstrates the need for carefully choosing the culture media and supplements due to the effect of steroid hormones.

Key words: phenol red, steroid hormones, CFTR, endometrial epithelial cells, short-circuit current.

^f1To whom correspondence should be addressed: Prof. H. C. Chan, Department of Physiology, The Chinese University of Hong Kong, Shatin, Hong Kong, Tel: (852) 2609 6839 Fax: (852) 2603 5022; E-mail:hsiaocchan@cuhk.edu.hk

Received 9 January 2001; accepted 13 February 2001