Establishment of hypoxic conditions for cultured monolayer cells: hypoxia in five minutes or less.
A versatile deaeration chamber for altering the gaseous environment of monolayer cell cultures has been developed. The design is such that multiple samples of radioactively labeled or unlabeled cells can be gassed simultaneously with a short period of time. Deaeration times to reduce the level of oxygen in initially aerated cellular medium to very low levels (less than 20 parts per million) were decreased at least 6- to 20-fold when compared to most other methods commonly used to achieve N2- or N2O-induced hypoxia. The application of this chamber in the investigation of radiation-induced cellular DNA strand breaks, which are very sensitive to the presence of oxygen, is described. The vessel should be useful for a variety of metabolic and physiologic experiments or cell, tumor, and radiation biology studies.
Laboratory of Radiobiology, Harvard University School of Public Health, Boston, Massachusetts 02134.
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D015687:Cell Hypoxia; D002478:Cells, Cultured; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D005347:Fibroblasts / Q000166:cytology / Q000528:radiation effects; D005898:Glass; D006801:Humans; D010100:Oxygen / Q000502:physiology; D011853:Radiobiology / Q000379:methods
D004247:DNA; D010100:Oxygen