Reznik V M, Villela J, Mendoza S A
J Cell Physiol. 1983 Nov;117(2):211-4. doi: 10.1002/jcp.1041170212.
The growth of an epithelial canine kidney line (MDCK) was reversibly arrested by gradually lowering the serum concentration in the medium over a 3-day period. The cells were demonstrably quiescent by autoradiography after an additional 24 hours in serum-free media. Addition of fresh serum produced DNA synthesis after an 18-hour lag period. The quiescent cells then grew to confluency retaining their transport capacities as seen by the formation of "domes." This system allows for measurement of monovalent ion fluxes and its relationship to growth regulation. The addition of fresh serum to quiescent MDCK cells increased the uptake of 86Rb, a measure of Na-K pump activity. This stimulation was mediated by increased uptake of Na into the cells. Serum-stimulated DNA synthesis was blocked by the addition of ouabain in concentrations that inhibit the Na-K pump. Serum appears to stimulate growth in epithelial cells by increasing the amount of intracellular Na available to the Na-K pump. Monovalent ion transport may play a role in the regulation of epithelial cell proliferation.
通过在3天时间内逐渐降低培养基中的血清浓度,可使犬肾上皮细胞系(MDCK)的生长可逆性停滞。在无血清培养基中再培养24小时后,通过放射自显影法可明显看出细胞处于静止状态。添加新鲜血清后,经过18小时的延迟期会产生DNA合成。静止细胞随后生长至汇合状态,并保持其运输能力,这可通过“穹顶”的形成看出。该系统可用于测量单价离子通量及其与生长调节的关系。向静止的MDCK细胞中添加新鲜血清会增加86Rb的摄取,这是钠钾泵活性的一种测量指标。这种刺激是由细胞对钠摄取的增加介导的。添加抑制钠钾泵的哇巴因可阻断血清刺激的DNA合成。血清似乎通过增加钠钾泵可利用的细胞内钠量来刺激上皮细胞生长。单价离子运输可能在上皮细胞增殖的调节中起作用。