Extracellular acidosis delays cell death against glucose-oxygen deprivation in neuroblastoma x glioma hybrid cells

Critical Care Medicine
Y MorimotoE S Alojado

Abstract

To determine whether extracellular acidosis delays cell death against glucose-oxygen deprivation and, if so, whether this result is due to inhibition of calcium (Ca2+) influx or preservation of cellular energy state. Randomized, controlled, prospective study. University research laboratory. Differentiated neuroblastoma x glioma NG108-15 cells. Experiment 1: cells were incubated for 8 hrs in N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid-buffered medium under glucose-oxygen deprivation at pH 7.4, 6.8, 6.5, 6.2, 5.6, or 5.0. Experiment 2: cells were incubated for 8 hrs under glucose-oxygen deprivation after excluding extracellular calcium from culture medium at pH 7.4 or 6.2. Experiment 3: cells were incubated for 2, 4, 6, or 8 hrs in N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid-buffered medium under glucose-oxygen deprivation at pH 7.4 or 6.2 and assayed for high-energy phosphates. Cell viability was measured with flow cytometry after the cells were stained with fluorescein diacetate and propidium iodide. Cellular adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate were analyzed with high-performance liquid chromatography. Cell viability was significantly greater at pH 6.2 than at pH 7.4 in...Continue Reading

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