Detection of Apoptosis Induced by Ethanol in Hybridoma Cells Culture

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.