Walsh-Reitz M M, Toback F G
Department of Medicine, University of Chicago, Illinois 60637.
Am J Physiol. 1992 Apr;262(4 Pt 2):F687-91. doi: 10.1152/ajprenal.1992.262.4.F687.
Phenol red coeluted with a novel kidney cell growth factor during its isolation from conditioned medium by high-performance liquid chromatography. The possibility that phenol red rather than the putative factor mediated the growth-promoting activity was tested, because this pH indicator is known to possess estrogenic properties including a mitogenic effect. Unexpectedly, phenol red at the concentration found in Dulbecco's modified Eagle's medium (DMEM) inhibited growth of monkey (BSC-1) and canine kidney (MDCK) epithelial cells by 20-30%, but stimulated multiplication of 3T3 fibroblasts. The growth-inhibitory effect of phenol red for BSC-1 cells was reversible, concentration dependent, and cell-density independent when multiplication was examined in the presence and absence of the dye. Phenol red partially masked the mitogenic effect of calf serum and epidermal growth factor and was additive to the growth-inhibitory effect of transforming growth factor-beta 2. These observations indicate that phenol red at the concentration in DMEM both underestimates the potency of mitogens and overestimates the strength of an inhibitor of kidney epithelial cell growth and suggest that the dye be omitted from the culture medium when a new growth-regulatory compound is under study.
在通过高效液相色谱从条件培养基中分离一种新型肾细胞生长因子的过程中,酚红与之共洗脱。由于已知这种pH指示剂具有包括促有丝分裂作用在内的雌激素特性,因此对是酚红而非假定因子介导生长促进活性的可能性进行了测试。出乎意料的是,在杜氏改良伊格尔培养基(DMEM)中发现的浓度的酚红抑制了猴(BSC - 1)和犬肾(MDCK)上皮细胞的生长20 - 30%,但刺激了3T3成纤维细胞的增殖。当在有和没有该染料的情况下检测增殖时,酚红对BSC - 1细胞的生长抑制作用是可逆的、浓度依赖性的且与细胞密度无关。酚红部分掩盖了小牛血清和表皮生长因子促有丝分裂的作用,并且与转化生长因子-β2的生长抑制作用相加。这些观察结果表明,DMEM中浓度的酚红既低估了有丝分裂原的效力,又高估了肾上皮细胞生长抑制剂的强度,并表明在研究一种新的生长调节化合物时,应从培养基中省略该染料。