PH CHANGES IN PULSED CO2 INCUBATORS CAUSE PERIODIC CHANGES IN CELL MORPHOLOGY
PH CHANGES IN PULSED CO2 INCUBATORS CAUSE PERIODIC CHANGES IN CELL MORPHOLOGY
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DOI：
10.1006/excr.1994.1214
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发表时间：
1994-08-01
影响因子：
3.7
通讯作者：
GIAEVER, I
中科院分区：
文献类型：
--
作者：
LO, CM;KEESE, CR;GIAEVER, I
在动物细胞培养中，通常使用用重碳酸盐离子缓冲的培养基，并提高培养箱气体中的二氧化碳浓度。许多现代孵化器通过监测二氧化碳水平并在值降至特定设定点以下时注入气体来维持这些浓度。该反馈系统导致孵化器中二氧化碳浓度约为0.5%的微小振荡，从而导致沐浴细胞的介质中的pH值振荡。这些振荡非常小，使用标准的pH组合电极无法检测到，到目前为止，还没有人注意到对细胞的任何影响。然而，在这项研究中，我们证明了这些周期性的变化可以导致细胞形态的可检测变化，从而确实影响细胞的行为。使用电池-衬底阻抗传感(ECIS)检测细胞形态的细微变化，我们测量了细胞变化，当使用快速傅立叶变换分析时，这些变化具有与二氧化碳添加相匹配的周期性。当培养基因细胞新陈代谢而酸化，降低了液体的缓冲能力时，这种影响就变得特别明显。在ECIS测量中，我们使用细胞-电极相互作用模型进一步分析了周期性的形态变化。我们的结论是：(1)细胞形态的周期性变化在培养液部分耗尽和培养血管表面积与体积比高的情况下最为明显；(2)对于融合的WI-38/VA13成纤维细胞和牛肺内皮细胞(B3B3)，主要的周期性形态变化主要表现为细胞旁阻力的变化。(C)1994年学术出版社。
It is common in animal cell culture to utilize media that are buffered with bicarbonate ions and elevated CO2 concentrations in the incubator gas. Many modern incubators maintain these concentrations by monitoring the level of CO2 and injecting the gas when the value drops below a certain set point. This feedback system results in small oscillations in the incubator CO2 concentration of about 0.5%, resulting in oscillating pH values in the medium bathing the cells. These oscillations are very small and cannot be detected using a standard pH combination electrode, and until now no one has noticed any effect on the cells. In this study, however, we demonstrate that these periodic changes can result in detectable changes in cell morphology and therefore do affect cell behavior. Using electric cell-substrate impedance sensing (ECIS) to detect subtle alterations in cell morphology, we have measured cellular changes that when analyzed with the Fast Fourier Transform have a periodicity that matches those of the CO2 addition. This effect becomes particularly prominent when the culture medium has been acidified by cell metabolism reducing the liquid's buffer capacity. We have further analyzed the cyclical morphological changes employing a model of cell-electrode interactions in the ECIS measurement. We conclude that: (1) the periodic changes in cell morphology are most apparent if the medium is partially spent and if the surface to volume ratio of the culture vessel is high and (2) for confluent WI-38/VA13 fibroblasts and bovine pulmonary endothelial cells (B3B3), the main periodic morphological changes manifest themselves primarily as alterations in paracellular resistance. (C) 1994 Academic Press, Inc.