ELECTRON-SPIN-RESONANCE STUDIES OF O2 UPTAKE BY CHINESE-HAMSTER OVARY CELLS DURING THE CELL-CYCLE
ELECTRON-SPIN-RESONANCE STUDIES OF O2 UPTAKE BY CHINESE-HAMSTER OVARY CELLS DURING THE CELL-CYCLE
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DOI：
10.1073/pnas.79.4.1166
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
LUKIEWICZ, S
中科院分区：
文献类型：
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作者：
LAI, CS;HOPWOOD, LE;LUKIEWICZ, S
溶解的O2和氮氧自由基自旋探针之间的磁相互作用导致的ESR线加宽。一个封闭的室的方法基于此属性被用来确定在培养的哺乳动物细胞中的最大速率的O2摄取每细胞（Vmax每细胞）。将合适的自旋探针和细胞悬浮液在充气介质中混合，并测量溶解的O2的消失速率。研究了温度、pH值和微波功率对溶解氧测定的影响。对于异步中国仓鼠卵巢细胞，O2摄取为3.8 × 10 - 4。107个O2分子/细胞/s，并且在O2浓度> 10 μ M时似乎受到酶促限制。约5-10倍。每次测量使用105个细胞，使得有可能研究有丝分裂同步的细胞。使用这种方法，Vmax每单位细胞体积的变化，在中国仓鼠卵巢细胞的细胞周期中，从有丝分裂的最小值到G1和后期S期的最大值。自旋探针的优点和局限性进行了讨论，用于研究完整细胞的O2摄取。
Magnetic interactions between dissolved O2 and nitroxide radical spin probes lead to broadening of the ESR lines. A closed-chamber method based on this property was used to determine the maximum rate of O2 uptake per cell (Vmax per cell) in cultured mammalian cells. A suitable spin probe and a cell suspension are mixed in an aerated medium, and the rate of disappearance of dissolved O2 is measured. The effects of temperature, pH and microwave power on the determination of disolved O2 in solution were studied. For asynchronous Chinese hamster ovary cells, O2 uptake is 3.8 .times. 107 O2 molecules per cell per s and appears to be enzymatically limited at O2 concentrations > 10 .mu.M. About 5-10 .times. 105 cells were used for each measurement, making it possible to study mitotically synchronized cells. Using this method, Vmax per unit cell volume changes during the cell cycle of Chinese hamster ovary cells from a minimum in mitosis to maxima in both G1 and late S phases. Advantages and limitations of spin probes for studying the O2 uptake of intact cells are discussed.