乙醇诱导杂交瘤细胞凋亡的检测

为建立杂交瘤细胞凋亡检测模型，在乙醇诱导下，采用荧光染色、MTT等方法检测H18杂交瘤细胞凋亡时的形态学及增殖活性变化，并用流式细胞仪对其进行定量分析，夹心ELISA检测IgG抗体分泌的变化情况。结果发现5lOmmol／L乙醇作用5～6h的凋亡诱导效果最为明显，在诱导条件下，杂交瘤细胞的活细胞数显著下降，凋亡细胞比例较高。抗体浓度明显下降，与乙醇浓度呈负相关，但与细胞增殖活性无明显的线性关系。故以此为凋亡检测模型，可为后续抗凋亡细胞株的建立与筛选研究提供初步的实验基础，并为乙醇对IgG型抗体分泌的影响研究提供了可能的实验模型。

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.