Gabridge M G, Polisky R B
In Vitro. 1977 Aug;13(8):510-6. doi: 10.1007/BF02615144.
The amount of adenosine triphosphate (ATP) in hamster trachea organ cultures was determined with a technique based on light emission from a luciferin/luciferase/ATP reaction. The amount of ATP, expressed as ng per mg dry weight, was consistent in tracheal explants prepared from various animals and changed negligibly when explants were cultivated in vitro for several days. The amount of ATP was related directly to cellular activity and integrity in the epithelium since inactivation by heat or freeze-thaw rapidly depleted measurable ATP, and ciliary activity and ATP content were related directly. When tracheal explants were infected with 10(5) to 10(7) CFU of virulent Mycoplasma pneumoniae cells, both ciliary activity and ATP content in the tissue dropped dramatically after approximately 5 to 8 days (up to 85% and 60% decreases, respectively). Exposure of explants to 50 to 200 microgram per ml of purified M. pneumoniae membranes also caused significant decreases in ciliary activity and ATP. When explants were infected with attenuated or nonvirulent mycoplasmas, ciliary activity was only slightly decreased, while ATP values often rose slightly. The technology associated with the determination of ATP levels in tracheal explants should prove useful as a new, objective, analytical approach to cell viability in organ cultures.
采用基于荧光素/荧光素酶/ATP反应发光的技术，测定了仓鼠气管器官培养物中三磷酸腺苷（ATP）的含量。以每毫克干重中ATP的纳克数表示，从不同动物制备的气管外植体中的ATP含量是一致的，并且当外植体在体外培养数天时，ATP含量的变化可以忽略不计。由于热灭活或冻融能迅速消耗可测量的ATP，所以ATP的含量与上皮细胞的活性和完整性直接相关，而且纤毛活性和ATP含量也直接相关。当气管外植体感染10⁵至10⁷CFU的强毒肺炎支原体细胞时，大约5至8天后，组织中的纤毛活性和ATP含量均显著下降（分别下降高达85%和60%）。将外植体暴露于每毫升50至200微克的纯化肺炎支原体膜中，也会导致纤毛活性和ATP显著下降。当外植体感染减毒或无毒支原体时，纤毛活性仅略有下降，而ATP值通常略有上升。与测定气管外植体中ATP水平相关的技术，应可证明是一种用于评估器官培养中细胞活力的新的、客观的分析方法。
