Flow cytometric methods of analyzing apoptotic cells.
Flow cytometric methods of analyzing apoptotic cells.
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DOI：
10.1007/978-1-59259-257-9_35
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发表时间：
1998
影响因子：
--
通讯作者：
A. Milner;J. Michaela Levens;Christopher D. Gregory
中科院分区：
文献类型：
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作者：
A. Milner;J. Michaela Levens;Christopher D. Gregory
细胞凋亡是一种生理的、程序化的细胞死亡模式，对于胚胎发育中的组织建模和器官发生以及控制各种组织类型的动态平衡是必要的。细胞凋亡的独特形态特征清楚地将其与被动的细胞死亡、坏死模式区分开来，后者是对有毒刺激的一种非程序性反应。细胞凋亡的诊断依赖于对这些形态变化的检测，即染色质凝聚和碎裂，与细胞质凝聚相关的细胞收缩，以及细胞膜的完整性保留。这些变化可能伴随着细胞分裂成包含细胞质细胞器、核成分或两者的膜结合的凋亡体(2)。经典的细胞凋亡诊断方法包括光镜和电子显微镜，可显示上述形态变化；核小体间DNA片段化分析，凝胶电泳法可见DNA片段化，呈梯状，代表多个寡核体片断的产生。随着体外系统在细胞死亡研究中的广泛应用，许多研究人员转而使用流式细胞术来替代传统的分析细胞凋亡的方法，因为它具有对单个细胞的分析速度快、细胞数量要求低以及可以同时测量多个细胞参数的优点。
Apoptosis is a physiological, programmed mode of cell death, which is necessary for tissue modeling and organogenesis in embryonic development and in the control of homeostasis in a diversity of tissue types (1). The distinct morphological features of apoptosis clearly distinguish it from the passive mode of cell death, necrosis, which is an unprogrammed response to toxic stimuli. The diagnosis of apoptosis relies on detection of these morphological changes, i.e., condensation and fragmentation of chromatin, cell shrinkage associated with cytoplasmic condensation, and retention of cell membrane integrity. These changes may be accompanied by the fragmentation of the cell into membrane-bound apoptotic bodies containing cytoplasmic organelles, nuclear components, or both (2). Classical methods of diagnosing apoptosis include light and electron microscopy in which the above morphological changes can be visualized, and internucleosomal DNA fragmentation assays in which the fragmentation of DNA can be seen after gel electrophoresis as a ladder pattern representing the generation of multiple oligonucleosomal fragments. The widespread use of in vitro systems for studying cell death has led many researchers to turn to flow cytometry as an alternative to the classic methods of analyzing apoptosis, since it offers the advantages of rapid analysis of individual cells, low cell number requirement, and the opportunity for simultaneous measurement of several cellular parameters.