A method for measuring the oxygen consumption of intact cell monolayers
A method for measuring the oxygen consumption of intact cell monolayers
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DOI：
10.1152/ajplung.2000.278.4.l858
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发表时间：
2000-04-01
影响因子：
4.9
通讯作者：
Saumon, G
中科院分区：
文献类型：
--
作者：
Mamchaoui, K;Saumon, G
本报告描述了一种测量完整单层培养细胞的 O(2) 消耗 (Q) 点 O(2)) 的露天方法。该方法基于菲克第二扩散定律。它只需要一个显微操作器和一个微型 O(2) 电极即可测量孔中培养基中的 Po(2) 梯度。与传统的氧描记室法进行了比较。在新鲜分离的大鼠 II 型细胞中，两种方法给出的 (Q) 相对于点 O(2) 的值相同：166 +/- 15.3 nmol。 h(-1)。露天方法为 10(6) 个细胞(-1)，151 +/- 11.6 nmol。 h(-1)。 10(6) 个细胞(-1) 用于氧描记室方法（n = 11 次实验）。但在培养 2 天的细胞中（236 +/- 8.8 nmol . h(-1). 10(6) 个细胞(-1)），露天方法给出的 (Q) 值显着大于点 O(2)，而氧描记法 (71 +/- 15.2 nmol . h(-1). 10(6) 个细胞(-1)；P < 0.001；n = 12 次实验）。这表明细胞从基质上分离并放入氧描记室的方式会影响它们的 Q (2)。露天方法可能对研究单层细胞的代谢特性有用，因为细胞没有受损的风险。
This report describes an open-air method for measuring the O(2) consumption (Q) over dot O(2)) of intact monolayers of cultured cells. This method is based on Fick's second law of diffusion. It requires only a micromanipulator and a miniature O(2) electrode to measure the Po(2), gradient in the culture medium in the well. It was compared with the conventional oxygraph chamber method. Both methods gave the same value for (Q) over dot O(2) in freshly isolated rat type II cells: 166 +/- 15.3 nmol . h(-1). 10(6) cells(-1) for the open-air method and 151 +/- 11.6 nmol . h(-1). 10(6) cells(-1) for the oxygraph chamber method (n = 11 experiments). But the open-air method gave significantly larger values for (Q) over dot O(2) in cells cultured for 2 days (236 +/- 8.8 nmol . h(-1). 10(6) cells(-1)) than the oxygraph method (71 +/- 15.2 nmol . h(-1). 10(6) cells(-1); P < 0.001; n = 12 experiments). This suggests that the way cells are detached from their substratum to be placed in the oxygraph chamber affects their Q (2). The open-air method may be useful for studies on the metabolic properties of monolayers because the cells do not risk being damaged.