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使用流式细胞术检测异质细胞群体中的细胞凋亡。

Detection of apoptosis in a heterogenous cell population using flow cytometry.

作者信息

Sedlák J, Hunáková L, Duraj J, Suliková M, Chovancová J, Novotný L, Chorváth B

机构信息

Department of Molecular Immunology, Cancer Research Institute, Slovak Academy of Sciences, Bratislava.

出版信息

Gen Physiol Biophys. 1999 Oct;18 Spec No:147-54.

Abstract

Apoptosis induced in human leukemic cells (promyelocytic human leukemic cells HL-60, multidrug-resistant subline HL-60/VCR) and human ovarian carcinoma cells (A2780 and multidrug-resistant subline A2780/ADR) in vitro was detected by flow cytometric analysis or DNA electrophoresis. The cytofluorometric techniques utilized, i. e. detection of phosphatidylserine exposed at the outer surface of the plasma membrane, identification of cells with "sub-G0" DNA content or increased light side scatter (cell internal structure) correlated with the electrophoretic determination of DNA fragmentation ("DNA ladder"). Detection of the 34 kDa mitochondrial protein recognized by the monoclonal antibody Apo2.7 yielded elevated percentages of apoptotic cells, suggesting that this technique detecting both early and late apoptosis in digitonin-fixed cells might not be restricted to the specific detection of programmed cell death.

摘要

通过流式细胞术分析或DNA电泳检测体外诱导人白血病细胞(早幼粒细胞白血病细胞HL - 60、多药耐药亚系HL - 60/VCR)和人卵巢癌细胞(A2780和多药耐药亚系A2780/ADR)凋亡的情况。所采用的细胞荧光技术,即检测暴露于质膜外表面的磷脂酰丝氨酸、鉴定具有“亚G0”DNA含量的细胞或增加的侧向光散射(细胞内部结构),与DNA片段化的电泳测定(“DNA梯带”)相关。用单克隆抗体Apo2.7识别的34 kDa线粒体蛋白检测显示凋亡细胞百分比升高,这表明这种在洋地黄皂苷固定细胞中检测早期和晚期凋亡的技术可能不限于程序性细胞死亡的特异性检测。

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