Unveiling the DNA repair landscape by unlocking the potential of cryopreserved PBMCs with optimal processing

In the field of DNA repair, cryopreserved peripheral blood mononuclear cells (PBMCs) are a feasible model for large-scale human biomonitoring studies. However, the optimal processing of cryopreserved PBMCs to accurately represent real-time and reproducible DNA repair capacity (DRC) for an individual remains uncertain. The literature is inconclusive regarding the preservation of DRC among PBMC subpopulations or whether a particular cell type is a better representative of DRC. Additionally, it is unknown whether an individual’s DRC remains stable over time. This paper utilizes a functional-based Fluorescent Multiplex Host Cell Reactivation assay to evaluate DRC by measuring repair capability of intact PBMCs for various DNA lesions. The results demonstrate significant differences in DRC among PBMC subtypes during resting and stimulated stages, with T cells dominating most of the data obtained from PBMCs in all functional assays examined. The stability of DRC within individuals is maintained for major DNA repair pathways across repeated measurements. Furthermore, the rank order of individual DRC is preserved by transfecting cryopreserved resting human PBMCs. These findings have the potential to revolutionize approaches for handling cryopreserved PBMCs in the field of DNA repair for human bio-monitoring purposes.