Sioen Simon, Cloet Karlien, Vral Anne, Baeyens Ans
Radiobiology, Department of Human Structure and Repair, Ghent University, Corneel Heymanslaan 10, 9000 Gent, Belgium.
J Pers Med. 2020 Sep 14;10(3):125. doi: 10.3390/jpm10030125.
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 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.
胞质分裂阻滞微核（CBMN）试验是一种用于遗传毒性研究的标准化方法。虽然该试验也可在分离的外周血单核细胞（PBMC）培养物上进行，但传统的全血培养（WBC）常用于此试验。然而，PBMC CBMN试验方案的标准化尚未得到广泛研究。本研究的目的是优化一种针对新鲜和冷冻保存的外周血单核细胞（PBMC）的可靠CBMN试验方案，并比较WBC和PBMC培养物之间的微核（MNi）结果。对来自健康人血样和五个放射敏感患者血样的全血、新鲜分离的和冷冻保存的PBMC进行了G CBMN试验。细胞在体外暴露于0.5至2 Gy范围内的220 kV X射线剂量。优化后的PBMC CBMN试验显示出足够的重复性和较小的个体间变异性。WBC的MNi值显著高于新鲜PBMC。此外，PBMC的冷冻保存导致MNi值显著增加，而不同的冷冻保存时间没有显著影响。总之，我们针对新鲜和冷冻保存的PBMC的标准化CBMN试验可用于遗传毒性研究、生物剂量测定和放射敏感性评估。
