Application of a modified culturing method for lymphocytes in cytogenetic research / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a modified method for microculturing whole human blood for cytogenetic analysis.</p><p><b>METHODS</b>A novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.</p><p><b>RESULTS</b>The SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.</p><p><b>CONCLUSION</b>A modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.</p>

Effects of indium on micronucleus formation in human peripheral blood lymphocytes / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the cytotoxicity of indium chloride (InCl₃) and its effects on micro-nucleus formation in primary human lymphocytes cultured in vitro.</p><p><b>METHODS</b>The CCK-8 assay was used to evaluate the cytotoxicity of 24 h exposure to different concentrations of InCl₃(4, 40, 80, 200, 500, and 1 000 µmol/L) in lymphocytes cultured in vitro. The cytokinesis-block method was used to determine the micronucleus level in lymphocytes exposed to different concentrations of InCl₃and the effects of anti-oxidant vitamin C on micronucleus frequency.</p><p><b>RESULTS</b>Lymphocytes exposed to InCl₃of no less than 500 µmol/L had significantly lower survival rates than those in the control group (P < 0.05). Lymphocytes exposed to 80 µmol/L InCl₃had a significantly higher micronucleus frequency than those in the control group (P < 0.05). However, there was no further increase in micronucleus frequency of lymphocytes exposed to 200 µmol/L InCl₃. Lymphocytes cultured in whole blood and exposed to 500 or 1000 µmol/L InCl₃had a significantly increased micronucleus frequency than those in the control group (P < 0.001). The increase in micronucleus frequency of lymphocytes induced by indium could be partially antagonized by 20 or 100 µmol/L vitamin C.</p><p><b>CONCLUSION</b>InCl₃can induce an increase in micronucleus frequency of primary human lymphocytes cultured in vitro, which might be associated with DNA damage induced by oxidative stress.</p>