乙醇诱导杂交瘤细胞凋亡的检测  

Detection of Apoptosis Induced by Ethanol in Hybridoma Cells Culture

作　　者：徐静[1] 王贤辉[1] 米力[1] 陈志南[1] 

机构地区：[1]第四军医大学细胞生物学教研室,西安710032

出　　处：《细胞生物学杂志》2004年第2期185-188,共4页Chinese Journal of Cell Biology

基　　金：国家高技术研究发展计划(863计划)重大专项(2002AA217011)~~

摘　　要：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510mmol/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。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.

关 键 词：乙醇 杂交瘤细胞 凋亡 细胞培养 

分 类 号：Q813[生物学—生物工程]