An in Vitro ¹²⁵IUdR-release Assay for Measuring the Kinetics of Cell Death

Author： Schneiderman M.H. Hofer K.G. Schneiderman G.S.

ISSN： 1362-3095

Source： International Journal of Radiation Biology, Vol.59, Iss.2, 1991-02, pp. : 397-408

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Abstract

A radionuclide release assay for measuring the in vitro kinetics of cell death has been developed. CHO cells were labelled for 24 h with 3·0 hBq/ml of [¹²⁵I] iododeoxyuridine (¹²⁵IUdR) and the fate of the labelled cells and their progeny was monitored at daily intervals by measuring the rate of ¹²⁵I release. Prelabelling with ¹²⁵IUdR did not alter the plating efficiency, the doubling time or the selection of mitotic cells. The rate of ¹²⁵I release from labelled (but otherwise untreated) CHO cells was ≈ 4% day. Treatment with a lethal dose of X-rays (30 Gy), heat (46°C, 1 h), cold (− 90°C, 1 h) or the antibiotic Geneticin® (300 μg/ml, continuously) resulted in the release of > 99% the ¹²⁵I activity associated with the cells. Cell death was rapid after heating or freezing, and delayed after treatment with X-rays or Geneticin. The results illustrate the efficacy of the ¹²⁵I release assay for measuring the kinetics of cell death in mammalian tissue culture cells.