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The cytokinesis-block micronucleus assay on human isolated fresh and cryopreserved peripheral blood mononuclear cells

Simon Sioen (UGent) , Karlien Cloet, Anne Vral (UGent) and Ans Baeyens (UGent)
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Organization
Abstract
The cytokinesis-block micronucleus (CBMN) assay is a standardized method used for genotoxicity studies. Conventional whole blood cultures (WBC) are often used for this assay, although the assay can also be performed on isolated peripheral blood mononuclear cell (PBMC) cultures. However, the standardization of a protocol for the PBMC CBMN assay has not been investigated extensively. The aim of this study was to optimize a reliable CBMN assay protocol for fresh and cryopreserved peripheral blood mononuclear cells (PBMCS), and to compare micronuclei (MNi) results between WBC and PBMC cultures. The G(0)CBMN assay was performed on whole blood, freshly isolated, and cryopreserved PBMCS from healthy human blood samples and five radiosensitive patient samples. Cells were exposed to 220 kV X-ray in vitro doses ranging from 0.5 to 2 Gy. The optimized PBMC CBMN assay showed adequate repeatability and small inter-individual variability. MNi values were significantly higher for WBC than for fresh PBMCS. Additionally, cryopreservation of PBMCS resulted in a significant increase of MNi values, while different cryopreservation times had no significant impact. In conclusion, our standardized CBMN assay on fresh and cryopreserved PBMCS can be used for genotoxicity studies, biological dosimetry, and radiosensitivity assessment.
Keywords
PBMCS, micronucleus assay, biological dosimetry, human blood, genotoxicity tests, radiosensitivity, WHOLE-BLOOD, CHROMOSOMAL RADIOSENSITIVITY, ISOLATED LYMPHOCYTES, COMET ASSAY, DNA-DAMAGE, CANCER, MICRONUTRIENTS, FREQUENCY, GAMMA, IL-2

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MLA
Sioen, Simon, et al. “The Cytokinesis-Block Micronucleus Assay on Human Isolated Fresh and Cryopreserved Peripheral Blood Mononuclear Cells.” JOURNAL OF PERSONALIZED MEDICINE, vol. 10, no. 3, 2020, doi:10.3390/jpm10030125.
APA
Sioen, S., Cloet, K., Vral, A., & Baeyens, A. (2020). The cytokinesis-block micronucleus assay on human isolated fresh and cryopreserved peripheral blood mononuclear cells. JOURNAL OF PERSONALIZED MEDICINE, 10(3). https://doi.org/10.3390/jpm10030125
Chicago author-date
Sioen, Simon, Karlien Cloet, Anne Vral, and Ans Baeyens. 2020. “The Cytokinesis-Block Micronucleus Assay on Human Isolated Fresh and Cryopreserved Peripheral Blood Mononuclear Cells.” JOURNAL OF PERSONALIZED MEDICINE 10 (3). https://doi.org/10.3390/jpm10030125.
Chicago author-date (all authors)
Sioen, Simon, Karlien Cloet, Anne Vral, and Ans Baeyens. 2020. “The Cytokinesis-Block Micronucleus Assay on Human Isolated Fresh and Cryopreserved Peripheral Blood Mononuclear Cells.” JOURNAL OF PERSONALIZED MEDICINE 10 (3). doi:10.3390/jpm10030125.
Vancouver
1.
Sioen S, Cloet K, Vral A, Baeyens A. The cytokinesis-block micronucleus assay on human isolated fresh and cryopreserved peripheral blood mononuclear cells. JOURNAL OF PERSONALIZED MEDICINE. 2020;10(3).
IEEE
[1]
S. Sioen, K. Cloet, A. Vral, and A. Baeyens, “The cytokinesis-block micronucleus assay on human isolated fresh and cryopreserved peripheral blood mononuclear cells,” JOURNAL OF PERSONALIZED MEDICINE, vol. 10, no. 3, 2020.
@article{8686319,
  abstract     = {The cytokinesis-block micronucleus (CBMN) assay is a standardized method used for genotoxicity studies. Conventional whole blood cultures (WBC) are often used for this assay, although the assay can also be performed on isolated peripheral blood mononuclear cell (PBMC) cultures. However, the standardization of a protocol for the PBMC CBMN assay has not been investigated extensively. The aim of this study was to optimize a reliable CBMN assay protocol for fresh and cryopreserved peripheral blood mononuclear cells (PBMCS), and to compare micronuclei (MNi) results between WBC and PBMC cultures. The G(0)CBMN assay was performed on whole blood, freshly isolated, and cryopreserved PBMCS from healthy human blood samples and five radiosensitive patient samples. Cells were exposed to 220 kV X-ray in vitro doses ranging from 0.5 to 2 Gy. The optimized PBMC CBMN assay showed adequate repeatability and small inter-individual variability. MNi values were significantly higher for WBC than for fresh PBMCS. Additionally, cryopreservation of PBMCS resulted in a significant increase of MNi values, while different cryopreservation times had no significant impact. In conclusion, our standardized CBMN assay on fresh and cryopreserved PBMCS can be used for genotoxicity studies, biological dosimetry, and radiosensitivity assessment.},
  articleno    = {125},
  author       = {Sioen, Simon and Cloet, Karlien and Vral, Anne and Baeyens, Ans},
  issn         = {2075-4426},
  journal      = {JOURNAL OF PERSONALIZED MEDICINE},
  keywords     = {PBMCS,micronucleus assay,biological dosimetry,human blood,genotoxicity tests,radiosensitivity,WHOLE-BLOOD,CHROMOSOMAL RADIOSENSITIVITY,ISOLATED LYMPHOCYTES,COMET ASSAY,DNA-DAMAGE,CANCER,MICRONUTRIENTS,FREQUENCY,GAMMA,IL-2},
  language     = {eng},
  number       = {3},
  pages        = {12},
  title        = {The cytokinesis-block micronucleus assay on human isolated fresh and cryopreserved peripheral blood mononuclear cells},
  url          = {http://dx.doi.org/10.3390/jpm10030125},
  volume       = {10},
  year         = {2020},
}

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