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Detection of Apoptosis Induced by Ethanol in Hybridoma Cells Culture
XU Jing, WANG Xian Hui, MI Li*, CHEN Zhi Nan
Department of Cell Biology, Fourth Military Medical University, Xi'an 710032, China
Abstract: To construct an apoptosis model, after induced by ethanol, the morphological changes of H18 hybridoma were observed by fluorescence microscope after Hoechst 33342-PI double fluorescence staining; the viability was examined by MTT assay; the proportion of apoptosis was assayed by flow cytometry; and the IgG antibody concentration was measured by sandwich enzyme-linked immunosorbent assay. The results showed that 510mmol/L ethanol led to the most evident effect of apoptosis after 5-6 hours induction and the viability of H18 hybridoma was notably decreased with peak apoptosis observed by flow cytometry. The antibody concentration was decreased with a negative correlation to the dose of ethanol, while with no linear relation to the viability of the detected cells. These results provided a detection model for the study on anti-apoptosis followed, while also provided a study model of the effect of ethanol on IgG secreted.