Figure 3

Measuring radiosensitivity. There are many assays for measuring radiosensitivity. The gold standard is a clonogenic assay where single cells are plated and allowed to grow for 1 to 4 weeks to assess ability to form colonies. (a) Colonies from fibroblasts cultured from a human skin sample. As it takes several weeks to culture fibroblasts and carry out a clonogenic assay, more rapid assays are often used. (b) An example of a more rapid assay is the G2 assay: a peripheral blood sample is taken, lymphocytes are stimulated to proliferate with the mitogen phytohemagglutinin, after 72 hours the cells are irradiated with 0.5 gray (Gy), and after 30 minutes colcemid is added for 60 minutes to arrest cells at metaphase that were in G2 when irradiated. The number of chromosome aberrations (arrows) is scored relative to unirradiated controls [41]. (c) Another example is the micronucleus assay: peripheral blood lymphocytes are irradiated with approximately 2 Gy and incubated for 2 days, cytochalasin B is added to prevent cytoplasm division after mitosis, and cells are harvested after 1 day and the number of micronuclei per 100 to 1,000 cells is scored [42]. Demonstration of cellular radiosensitivity in individuals with life-threatening radiotherapy toxicity or cancer-predisposing syndromes usually involves fibroblasts and derivation of radiation survival curves. (d) Survival curves for a number of individuals, including one (blue line) with ataxia telangiectasia, showing extreme cellular radiosensitivity. Parameters can be obtained from fitting curves to the data and parameters that reflect the initial slope, such as alpha and surviving fraction at 2 (SF2) or 3 Gy, are better at showing differences in radiosensitivity between people [133]. (e) Normal (that is, non-syndromic) individuals vary in radiosensitivity with a distribution that is approximately normal [134].