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

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 i...

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Bibliographic Details
Published in:International journal of radiation biology 2020-02, Vol.96 (2), p.214-219
Main Authors: Selvan Gnana Sekaran, Tamizh, Ricoul, Michelle, Brochard, Patricia, Herate, Cecile, Sabatier, Laure
Format: Article
Language:English
Subjects:
Adult
Akodon
Animals
Centromere - radiation effects
Centromere - ultrastructure
CHO Cells
Chromosomes - radiation effects
Chromosomes - ultrastructure
Cricetinae
Cricetulus
cytogenetics
Gamma Rays
Healthy Volunteers
Humans
lymphocytes
Lymphocytes - cytology
Middle Aged
Mitosis
premature chromosome condensation
Radiometry
Rodentia
Space life sciences
Telomere - radiation effects
Telomere - ultrastructure
Young Adult
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Summary: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 γ-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.
ISSN:0955-3002
1362-3095
DOI:10.1080/09553002.2019.1625493