Title: Establishment of flow cytometry assay of determination of cytotoxicity
Abstract: Objective:To establish a new method of measuring cell-mediated cytotoxicity by flow-cytometric assay.Methods:The test principle is based on target cell labeling with CFSE and subsequent DNA-labelling with PI for identification of target cells with compromised cell membranes.Effectors and targets were incubated at E/T ratios from 100∶1 to 6 25∶1 at different time points from 1 h to 6 h,collect 1 000 target events and analyze specific cytotoxicity.Results:CFSE was found to be a suitable fluorochrome for labelling targets cells since it showed a good separation between effectors and target cells;the proportion of spontaneously dead target cell is below 5%;the cytotoxic activity showed a significant and positive correlation with E/T ratio and incubation period;the optimal E/T ratio and incubation period is 50∶1~25∶1 and 2 h~4 h.Conclusion:The benefits of flow cytometry to measure cytotoxicity are as follows:avoid radioactive substance;increase sensitivity at early timepoint;analyse at cell-to-cell basis;long-term incubation period is allowed.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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