Quantitative assessment of markers for cell senescence
Quantitative assessment of markers for cell senescence
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DOI：
10.1016/j.exger.2010.01.018
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发表时间：
2010-10-01
影响因子：
3.9
通讯作者：
Passos, Joao F.
中科院分区：
文献类型：
--
作者：
Lawless, Conor;Wang, Chunfang;Passos, Joao F.
细胞衰老，复制能力的不可逆丧失，可能是肿瘤抑制因子，也是与年龄相关的组织功能丧失的原因之一。缺乏对衰老细胞的候选标记物的可靠性的定量测试是细胞群体研究中的主要缺点。培养中的成纤维细胞构成具有增殖能力的细胞和衰老细胞的混合群体，随着群体倍增（PD）的增加，这些细胞之间存在过渡。我们使用动态模拟模型从容易观察到的生长曲线以高精度估计人和小鼠成纤维细胞中的衰老分数。我们还确定了衰老分数，在不同的PD（在广泛的衰老细胞频率）使用候选衰老标记：Ki 67，p21（CDKN 1A），γ/H2 AX，SAHF和Sen-beta-Gal单独或组合，并与那些来自生长曲线。该比较允许对候选标记进行排序。对于Sen-beta-Gal、SAHF和Ki 67阴性与MRC 5成纤维细胞中的高（每个核>5个）γ H2A.X灶密度的组合，获得了高排名。我们证明，后者的标志物组合，这可以很容易地进行石蜡包埋的组织，在小鼠胚胎成纤维细胞培养物和小鼠肠道切片中的定量衰老细胞频率估计。提出的技术是一个定量评估衰老标记的框架。(C)2010年爱思唯尔公司All rights reserved.
Cellular senescence, the irreversible loss of replicative capacity, might be a tumour suppressor and a contributor to age-related loss of tissue function. The absence of quantitative tests for reliability of candidate markers for senescent cells is a major drawback in cell population studies. Fibroblasts in culture constitute mixed populations of proliferation-competent and senescent cells, with transition between these with increasing population doublings (PD). We estimated senescent fraction in human and mouse fibroblasts with high precision from easily observed growth curves using a dynamic simulation model. We also determined senescent fractions, at various PD (over a wide range of senescent cell frequencies) using candidate senescence markers: Ki67, p21 (CDKN1A), gamma/H2AX, SAHF and Sen-beta-Gal either alone or in combination, and compared with those derived from growth curves. This comparison allowed ranking of candidate markers. High rankings were obtained for Sen-beta-Gal, SAHFs and the combination of Ki67 negativity with high (>5 per nucleus) gamma H2A.X foci density in MRC5 fibroblasts. We demonstrate that this latter marker combination, which can easily be performed in paraffin-embedded tissue, gives quantitative senescent cell frequency estimates in mouse embryonic fibroblast cultures and in mouse intestinal sections. The technique presented is a framework for quantitative assessment of markers for senescence. (C) 2010 Elsevier Inc. All rights reserved.