Triage biodosimetry using centromeric/telomeric PNA probes and Giemsa staining to score dicentrics or excess fragments in non-stimulated lymphocyte prematurely condensed chromosomes

Dose assessment intercomparisons within the RENEB network using G0-lymphocyte prematurely condensed chromosomes (PCC assay)

“…Lymphocytes isolated from 1 ml of blood were used for 1–2 experimental points. Cell fusion and PCC induction using PEG has been described previously (Pantelias and Maillie, 1983, Vyas et al, 1991, Darroudi et al, 1998a, Karachristou et al, 2015, Sebastia et al, 2015). Briefly, mitotic CHO cells harvested from a 75 cm 2 flask were used for 2–3 fusions.…”

“…The supernatant was discarded and the cell pellet was resuspended gently in 0.7 ml RPMI-1640 complete growth medium with colcemid. To optimize cell fusion when a low number of lymphocytes is available, complete lymphocyte growth medium, containing 2% phytohemagglutinin (PHA) and 10% FBS, was used (Pantelias and Maillie, 1983, Karachristou et al, 2015, Sebastia et al, 2015). After 60–75 min at 37°C, cell fusion and PCC induction was completed.…”

“…In addition, Fomina and co-workers (Fomina et al, 2000, Fomina et al, 2001) developed and reported on a combination of staining all human centromeres and telomeres with chromosome paint probes in metaphases, in order to assess accurately different types of structural chromosomal aberrations in human lymphocytes, following exposure to radiation of different qualities. Recently, in order to score dicentric and centric ring chromosomes directly in G 0 -lymphocyte PCCs with a level of accuracy not possible previously, the development of centromeric/telomeric (C/T) peptide nucleic acid (PNA) probes in combination with FISH (C/T FISH staining), has offered a new dynamic to the PCC-assay (Karachristou et al, 2015, M’Kacher et al, 2015, Suto et al, 2015). …”

Uncertainty of fast biological radiation dose assessment for emergency response scenarios

“…However, it should be noted that recommendations differ between the assays. For the micronucleus assay, a larger number of approximately 200 cells is required (Thierens et al 2014); for gamma-H2AX and PCC, 20-30 cells may be sufficient Karachristou et al 2015). However, while uncertainties have been analyzed in order to provide an indication of the number of cells that might result in a suitably reliable dose estimate, the impact of uncertainty on the rapid biodosimetry categorization results has not been considered.…”

RENEB – Running the European Network of biological dosimetry and physical retrospective dosimetry

“…The methods included different types of the gene expression assay (qPCR, array based technology), thermoluminescence of mobile phone display glass, Raman spectroscopy and further developments of established methods as dicentric and PCC assay by combining them with centromere and telomere staining (M'kacher et al 2014, 2015; Karachristou et al 2015;Abend et al 2016). Some of these methods were included in RENEB intercomparisons and results are published in this special issue .…”