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Updated: Jun 22, 2026

Methods to Evaluate Cytotoxicity and Immunosuppression of Combustible Tobacco Product Preparations
Published on: January 10, 2015
[Cyto-genotoxicity induced by cigarette smoke condensates in human peripheral blood lymphocytes in vitro]
Jian-lin Lou1, Guo-jun Zhou, Guo-hai Chu
1Technical Center, China Tobacco Zhejiang Industrial Co., Ltd, Hangzhou 310008, China.
Objective:
To investigate the cyto-genotoxicity of cigarette smoke condensates (CSCs) in human peripheral blood lymphocytes with different assays in vitro.
Methods:
Human lymphocytes were exposed to particle matter of cigarette smoke combined with or without S9 mixtures at doses of 25, 50, 75, 100 and 125 microg/ml for 3 h. The cytotoxicity induced by CSCs was detected by CCK-8 assay. The DNA damage, DNA repair (repair time: 30, 60, 90, 120 and 240 min, respectively) and the somatic cell mutations induced by 75 microg/ml CSCs were measured by comet assay, hprt gene and TCR gene mutation tests, respectively.
Results:
CCK-8 assay indicated that the cell viability decreased with CSCs doses. At the doses of 100, 125 microg/ml, the cell viability of CSCs +S9 group was significantly higher than that of CSCs -S9 group (P < 0.05, P < 0.01). In comet assay, DNA damage significantly increased in a dose-dependent manner, as compared with controls (P < 0.01). Moreover, there was significant difference between -S9 group and +S9 group (P < 0.05, P < 0.01). The Mf-TCR at each dose group was significantly higher than that of controls (P < 0.05, P < 0.01). The Mf-hprt at high-dose groups were significantly higher than that of controls (P < 0.01), and significant difference of Mf-TCR and Mf-hprt at high doses of CSCs between -S9 group and +S9 group (P < 0.05, P < 0.01). The DNA damage induced by CSCs +S9 or CSCs -S9 could be repaired, but DNA repair speed was different between -S9 group and +S9 group (P < 0.05, P < 0.01).
Conclusion:
CSCs may induce cyto-genotoxicity in human peripheral blood lymphocytes in vitro, but S9 mix could reduce the toxicity of CSCs and impact DNA repair speed.
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