Neurology Department, Zhongshan Hospital, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai, China.
PLoS One. 2013;8(4):e60189. doi: 10.1371/journal.pone.0060189. Epub 2013 Apr 1.
Phenol red is widely used in cell culture as a pH indicator. Recently, it also has been reported to have estrogen-like bioactivity and be capable of promoting cell proliferation in different cell lines. However, the effect of phenol red on primary neuronal culture has never been investigated. By using patch clamp technique, we demonstrated that hippocampal pyramidal neurons cultured in neurobasal medium containing no phenol red had large depolarization-associated epileptiform bursting activities, which were rarely seen in neurons cultured in phenol red-containing medium. Further experiment data indicate that the suppressive effect of the phenol red on the abnormal epileptiform burst neuronal activities was U-shape dose related, with the most effective concentration at 28 µM. In addition, this concentration related inhibitory effect of phenol red on the epileptiform neuronal discharges was mimicked by 17-β-estradiol, an estrogen receptor agonist, and inhibited by ICI-182,780, an estrogen receptor antagonist. Our results suggest that estrogen receptor activation by phenol red in the culture medium prevents formation of abnormal, epileptiform burst activity. These studies highlight the importance of phenol red as estrogen receptor stimulator and cautions of careful use of phenol red in cell culture media.
酚红被广泛应用于细胞培养作为 pH 值指示剂。最近，它也被报道具有类雌激素生物活性，并能够促进不同细胞系的增殖。然而，酚红对原代神经元培养的影响尚未被研究过。通过使用膜片钳技术，我们证明了在不含酚红的神经基底培养基中培养的海马锥体神经元有大的去极化相关的癫痫样爆发活动，而在含有酚红的培养基中培养的神经元很少出现这种情况。进一步的实验数据表明，酚红对异常癫痫样爆发神经元活动的抑制作用呈 U 形剂量相关，最有效浓度为 28µM。此外，雌激素受体激动剂 17-β-雌二醇模拟了酚红对癫痫样神经元放电的这种浓度相关抑制作用，而雌激素受体拮抗剂 ICI-182,780 则抑制了这种作用。我们的结果表明，培养基中的酚红通过激活雌激素受体防止了异常癫痫样爆发活动的形成。这些研究强调了酚红作为雌激素受体激动剂的重要性，并提醒在细胞培养基中使用酚红时要小心谨慎。
