An alternative approach for the induction of premature chromosome condensation in human peripheral blood lymphocytes using mitotic Akodon cells

Abstract

Purpose: The premature chromosome condensation (PCC) technique is used to study exposure to external radiation through the determination of chromosome fragments observed in interphase cells. The presence of large telomeric signals in CHO cells interferes with the detection of PCC fragments and the identification of dicentric chromosomes. We present an improved method for the fusion of G0-lymphocytes with mitotic Akodon cells (few chromosomes and weakly-staining telomeric sequences) to induce PCC in combination with rapid quantification of dicentric chromosomes and centric rings as an alternative to the classical CHO cell fusion technique. Materials and methods: Whole blood from three healthy volunteers was c-irradiated with 0, 2, or 4 Gy. Following a 24 h incubation post-exposure at 37 C, chromosome spreads of isolated lymphocytes were prepared by standard PCC procedures using mitotic Akodon cells. Results: The percentage of scorable fusions, measured by telomere/centromere (T/C) staining, for Akodon-induced PCC was higher than that for CHO-induced PCC, irrespective of radiation exposure. Importantly, both techniques gave the same result for biodosimetry evaluation. Conclusion: The mitotic Akodon cell-induced PCC fusion assay, in combination with the scoring of dicentric chromosomes and rings by T/C staining of G0-lymphocytes is a suitable alternative for fast and reliable dose estimation after accidental radiation exposure. ARTICLE HISTORY

Cytogenetic biological dosimetry assays: recent developments and updates

“…Blind dose assessments were executed for triage biodosimetry investigating G0-lymphocyte PCC for standardization, harmonization, and validating the PCC assay [ 72 ]. It is also possible for automation [ 73 ], in addition, and new approaches have been introduced [ 74 ]. The combination of automatic image acquisition and automated image analysis of PCC allows calculating the length ratio of the longest and the shortest chromosome pieces that can be used for dose estimation [ 66 ].…”