Carcinogenesis, Teratogenesis & Mutagenesis ›› 2025, Vol. 37 ›› Issue (5): 400-404.doi: 10.3969/j.issn.1004-616x.2025.05.009

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Effect of whole blood inoculum and culture system on lymphocyte nucleoplasmic bridges formation

ZHAO Hua, LU Xue, CAI Tianjing, QI Xuesong, LIU Qingjie   

  1. China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China
  • Received:2025-05-21 Revised:2025-06-19 Published:2025-10-17

Abstract: OBJECTIVE: To investigate effects of whole blood inoculation and culture systems on formation of nucleoplasmic bridges (NPB) and on formation of radiation-induced NPB,for providing scientific basis for radiation biological dosimeters. METHODS:Human venous blood and fingertip blood were cultured as follows:400,300,200,100 and 50 μL of venous blood were inoculated into 1 or 2 mL medium,100 μL of fingertip blood were inoculated into 1 mL medium. These cultures were irradiated with 0 Gy (control group) and 2 Gy 60Co γ radiation at a dose rate of 1.0 Gy/min. NPB specimens were prepared by cytokinesis-block method. The proportion of mononucleated,binucleated and multinucleated cells,as well as the NPB and micronucleus (MN) frequencies of each group were analyzed. RESULTS:In the 0 Gy group, 1 048~9 768 binucleated cells were analyzed in different blood inoculation groups. The NPB frequency did not change significantly with the whole blood inoculation (P>0.05). The MN frequency in the 50 μL group was higher than that in the other three groups (U=3.61,P<0.01). In the 2 Gy group,there was no cell available for analysis in the 50 μL group,and 1 569-4 400 binucleated cells were analyzed in other blood inoculation groups. The NPB frequency was 0.009-0.013 cells/cell,and the MN frequency was 0.317-0.337 cells/cell,and there was no significant difference with the whole blood inoculum (P > 0.05). The NPB and MN frequencies of the 2 Gy group were significantly higher than those of the 0 Gy group (U=4.24-34.48, P<0.05). Fingertip blood culture failed to observe cells for analysis. CONCLUSION: The minimum 50 μL and 100 μL peripheral blood culture combined with 1 mL culture medium can be used to study the background level of the nucleoplasmic bridge and the radiation-induced nucleoplasmic bridge level respectively. The feasibility of fingertip blood culture for nucleoplasmic bridge analysis still needs further study.

Key words: nucleoplasmic bridge, whole blood inoculum, cytokinesis-block, micronucleus

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