Use of three-way differential staining and liquid holding for the assessment of individual repair capacity

Abstract

Many studies on DNA repair mechanisms in mammalian cells have used liquid holding (LH) recovery to evaluate premutational damage repair. We used human peripheral lymphocytes (HPL) to assess damage reduction during the G0 phase. This technique was matched with the three-way differential (TWD) staining that allows identification of sister-chromatid exchanges (SCE) per cell cycle in third metaphases. By adopting this approach, the persistence of diepoxybutane (DEB)-induced lesions during subsequent cycles and individual repair capacity in LH conditions were measured. Our results show that most DEB-induced damage was repaired during the first cell cycle; a large part of lesions were removed during LH recovery, demonstrating G0 HPL repair capacity.