A Simplified Calyculin A-Induced Premature Chromosome Condensation (PCC) Protocol for the Biodosimetric Analysis of High-Dose Exposure to Gamma Radiation

2020 ◽  
Vol 193 (6) ◽  
pp. 560 ◽  
Author(s):  
Mingzhu Sun ◽  
Jayne Moquet ◽  
Stephen Barnard ◽  
David Lloyd ◽  
Elizabeth Ainsbury
Keyword(s):  
Gamma Radiation ◽  
Chromosome Condensation ◽  
High Dose ◽  
Calyculin A ◽  
Premature Chromosome Condensation

Transient inhibition of Calyculin A induced premature chromosome condensation by hyperthermia

2009 ◽  
Vol 25 (3) ◽  
pp. 220-228
Author(s):  
J. W. J. Bergs ◽  
J. W. J. Bergs ◽  
R. Ten Cate ◽  
H. M. Rodermond ◽  
P. A. Jaarsma ◽  
...  
Keyword(s):  
Chromosome Condensation ◽  
Calyculin A ◽  
Premature Chromosome Condensation

scholarly journals Refined premature chromosome condensation (G0-PCC) with cryo-preserved mitotic cells for rapid radiation biodosimetry

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Usha Yadav ◽  
Nagesh N. Bhat ◽  
Kapil B. Shirsaath ◽  
Utkarsha S. Mungse ◽  
Balvinder K. Sapra
Keyword(s):  
Mitotic Index ◽  
Human Lymphocytes ◽  
Mitotic Cell ◽  
Chromosome Condensation ◽  
Blood Collection ◽  
High Dose ◽  
Premature Chromosome Condensation ◽  
Isolated Cells ◽  
Emergency Scenarios ◽  
Mitotic Cells

AbstractMitotic cell fusion induced Premature Chromosome Condensation (G0-PCC) assay in human lymphocytes allows rapid detection of cytogenetic damage in interphase stage, within few hours after blood collection. Hence, it is the most suitable method for rapid and high dose biodosimetry. Mitotic cells, used for G0-PCC could be either freshly isolated or previously cryo-preserved. However, under emergency scenarios, only cryo-preserved cells can be relied upon, fresh isolation will only delay the process by 18–24 h. Impact of cryopreservation on mitotic cells and their efficacy to induce PCC are not reported. In the present study, we investigated effect of cryopreservation on mitotic cells and refined the parameters for G0-PCC. More than 95% of the cells were recoverable after 4 months of cryopreservation, within 20 min recovery at 37 °C, without significant change in the mitotic index or viability. Recovered mitotic cells have shown mitotic index of 89 ± 4% and viability of 90 ± 4%, similar to that of freshly isolated cells. Decrease in metaphases was observed within 40 min after recovery as the mitotic cells progressed through cell cycle and reduced to 21% at 1.5 h. Nevertheless, in presence of Colcemid, the cells progressed slowly and considerably high metaphase index (60%) persisted up to ~ 2 h. The recovered cells efficiently fused with lymphocytes and induced PCC. Average PCC index varied from 10 to 20%, which did not change with cryopreservation duration. Post fusion incubation duration of 2 h was found to be optimum for proper chromosome condensation. In conclusion, use of cryo-preserved mitotic cells is the most practical approach for rapid biodosimetry. The cells can be recovered quickly and efficiently without alteration in viability or mitotic index. Recovered cells are fully competent to induce G0-PCC.

scholarly journals Detection of leukemic clone maturation in vivo by premature chromosome condensation

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 1950-1960
Author(s):  
WN Hittelman ◽  
P Agbor ◽  
I Petkovic ◽  
B Andersson ◽  
H Kantarjian ◽  
...  
Keyword(s):  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
High Dose Chemotherapy ◽  
Chromosome Condensation ◽  
Response To Therapy ◽  
Myelogenous Leukemia ◽  
High Dose ◽  
Premature Chromosome Condensation ◽  
Leukemic Clone

The purpose of this study was to determine the feasibility of using the technique of premature chromosome condensation to detect the in vivo maturation of abnormal elements in patients with chronic myelogenous leukemia (CML), myelodysplastic syndrome, and acute leukemia. Patients were chosen for study if there were a clinical suggestion of in vivo maturation and a leukemic clone exhibiting a distinguishable karyotypic abnormality. Mature peripheral blood granulocytes were enriched by two- step Ficoll-Hypaque gradient sedimentation and fused with mitotic Chinese hamster ovary cells to induce the formation of prematurely condensed chromosomes (PCC). These PCC were then analyzed for chromosome number per cell (in the case of patients with a numerical abnormality) or by G-banding (in the case of specific translocations). Of 13 patients chosen for study, 12 showed karyotypic evidence for maturation of the abnormal elements in vivo. Maturation was observed in a number of clinical situations including before treatment in benign CML and myelodysplasia, after low-dose and high-dose chemotherapy in myelodysplasia and acute myelogenous leukemia (AML), and in remission. These results suggest that the technique of premature chromosome condensation can be a powerful tool in better understanding the biology of disease and mode of response to therapy in vivo in patients with leukemia and preleukemic syndromes, especially during treatment with agents thought to induce maturation of the leukemic elements.

scholarly journals The Usefulness of Calyculin A for Cytogenetic Prenatal Diagnosis

2005 ◽  
Vol 53 (3) ◽  
pp. 391-394 ◽  
Author(s):  
Malgorzata I. Srebniak ◽  
Gizela G. Trapp ◽  
Angelika K. Wawrzkiewicz ◽  
Wojciech Kaźmierczak ◽  
Andrzej K. Wiczkowski
Keyword(s):  
Cell Cycle ◽  
Prenatal Diagnosis ◽  
Amniotic Fluid ◽  
Protein Phosphatases ◽  
Genetic Material ◽  
Chromosome Condensation ◽  
Calyculin A ◽  
Premature Chromosome Condensation ◽  
Chromosomal Breaks

An increased number of chromosome plates can be obtained by use of calyculin A (CLA). CLA is an inhibitor of protein phosphatases (type 1 and type 2A serine/threonine). Inactivation of these phosphatases leads to premature chromosome condensation (PCC) in all phases of the cell cycle; thus, it is possible to investigate both metaphase and G2-PCC chromosomes. Amniotic fluid (AF) cultures were treated with calyculin A (CLA). GTG banding was obtained. Using this method it is possible to investigate all cell cycle phases, GTG banding, chromosomal breaks, and rates of PCD on the same preparation. Analyses of AF cultures treated with CLA allow complex studies on fetal genetic material. This work presents potential usefulness of CLA for cytogenetic prenatal diagnosis.

scholarly journals Detection of leukemic clone maturation in vivo by premature chromosome condensation

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 1950-1960 ◽  
Author(s):  
WN Hittelman ◽  
P Agbor ◽  
I Petkovic ◽  
B Andersson ◽  
H Kantarjian ◽  
...  
Keyword(s):  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
High Dose Chemotherapy ◽  
Chromosome Condensation ◽  
Response To Therapy ◽  
Myelogenous Leukemia ◽  
High Dose ◽  
Premature Chromosome Condensation ◽  
Leukemic Clone

Abstract The purpose of this study was to determine the feasibility of using the technique of premature chromosome condensation to detect the in vivo maturation of abnormal elements in patients with chronic myelogenous leukemia (CML), myelodysplastic syndrome, and acute leukemia. Patients were chosen for study if there were a clinical suggestion of in vivo maturation and a leukemic clone exhibiting a distinguishable karyotypic abnormality. Mature peripheral blood granulocytes were enriched by two- step Ficoll-Hypaque gradient sedimentation and fused with mitotic Chinese hamster ovary cells to induce the formation of prematurely condensed chromosomes (PCC). These PCC were then analyzed for chromosome number per cell (in the case of patients with a numerical abnormality) or by G-banding (in the case of specific translocations). Of 13 patients chosen for study, 12 showed karyotypic evidence for maturation of the abnormal elements in vivo. Maturation was observed in a number of clinical situations including before treatment in benign CML and myelodysplasia, after low-dose and high-dose chemotherapy in myelodysplasia and acute myelogenous leukemia (AML), and in remission. These results suggest that the technique of premature chromosome condensation can be a powerful tool in better understanding the biology of disease and mode of response to therapy in vivo in patients with leukemia and preleukemic syndromes, especially during treatment with agents thought to induce maturation of the leukemic elements.

Sign in / Sign up

Export Citation Format

Share Document