G2 Chromosomal Radiosensitivity Assay for Testing Individual Radiation Sensitivity.

ABSTRACT

The G2 chromosomal radiosensitivity assay or, simply G2 assay, measures the number of chromatid type aberrations induced by radiation in G2 phase. Typically, asynchronous growing cells are irradiated with less than 1 Gy and allowed 0.5-1 h for cells in mitosis, at the time of irradiation, to transit into G1. Later, the G2 phase cells, at the time irradiation, are blocked by colcemid for 1-4 h at metaphase. Cells are collected by standard hypotonic solution and Carnoy solution fixation or directly fixed onto the culture vessels. The G2 assay can detect severe radiosensitivity in ATM homozygous mutated cells and relatively small differences among cellular radiosensitivity such as heterozygous mutation carriers of ATM and BRCA1/2 mutation carriers. The G2 assay also has the capability to detect cancer prone individuals. This assay only requires a conventional cell culture facility and the standard microscopic observation.