Differentiation and assessment of cell death
Differentiation and assessment of cell death
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DOI：
10.1515/cclm.1999.053
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发表时间：
1999-03-01
影响因子：
6.8
通讯作者：
Bolton, WE
中科院分区：
文献类型：
--
作者：
Koester, SK;Bolton, WE
三种记录的细胞死亡途径，凋亡，坏死和肿瘤将被讨论。每种途径的最终结果都是细胞死亡；然而，死亡的途径以及每种死亡的形态和生理特征可能是截然不同的。现在已经建立了表征良好的模型，特别是细胞凋亡，诱导通路受到了广泛的关注，通路病理也开始被理解。研究了三种模型系统：APO-1/Fas、缺氧和肿瘤。诱导细胞死亡，在时间过程取样过程中，采用流式细胞术和凝胶电泳DNA片段化、DNA染色、流式细胞术光散射、透射电镜、抗微管蛋白、泰潘蓝、膜联蛋白V、抗apo2.7等多种方法监测细胞死亡进程。利用caspase抑制剂肽进一步研究cd95诱导Jurkat细胞模型的凋亡途径，并利用流式细胞术分析APO2.7抗原表达和DNA片段化。时间过程采样表征了细胞死亡途径，并有助于区分方法的能力。反应的时间和持续时间取决于细胞类型和诱导方法。cd95诱导的Jurkat细胞模型表现出典型的凋亡反应；而MDA-MB-175-VII缺氧模型和抗5a9诱导的肿瘤模型则不明显。每种方法都显示出优点和缺点，使研究者能够选择几种方法来识别、监测和枚举细胞死亡进展，并使用时间过程研究。
Three documented cell death pathways, apoptosis, necrosis, and oncosis will be discussed. The end result of each pathway is cell death; however, the path by which death is achieved and the morphological and physiological traits of each may be strikingly distinct. Now that well characterized models have been established for particularly apoptosis, the induction pathway(s) has received much attention and the pathway pathology is beginning to be understood. Three model systems were investigated: APO-1/Fas, hypoxia, and oncosis. Cell death was induced, and during a time course sampling, a variety of methodologies, including DNA fragmentation by flow cytometry and gel electrophoresis, DNA staining, flow cytometric light scatter, transmission electron microscopy, anti-tubulin, Trypan blue, annexin V, and anti-APO2.7 were employed to monitor the cell death progress. The apoptotic pathway in the CD95-induced Jurkat cell model was further investigated using caspase inhibitor peptides and analyzed for APO2.7 antigen expression and DNA fragmentation by flow cytometry. Time course sampling characterized the cell death pathway and helped to differentiate the capabilities of the methods. The time to response and duration of the response were dependent upon cell type and method of induction. The CD95-induced Jurkat cell model showed a classical apoptotic response; however the MDA-MB-175-VII hypoxia model and the anti-5A9 induced oncosis model were not as clear. Each methodology shows advantages and disadvantages that allow the investigator to select several methods to identify, monitor, and enumerate cells with respect to cell death progression using time course studies.