Techniques to Induce and Quantify Cellular Senescence.
Techniques to Induce and Quantify Cellular Senescence.
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诱导和量化细胞衰老的技术。
DOI：
10.3791/55533
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发表时间：
2017-05-01
期刊：
影响因子：
--
通讯作者：
Evans MK
中科院分区：
文献类型：
--
作者：
Noren Hooten N;Evans MK
在应对细胞应激或损伤时，增殖细胞可以诱导一种特定的程序，启动长期细胞周期停滞的状态，称为细胞衰老。衰老细胞的积累是伴随着机体的衰老和体外的持续培养而发生的。衰老细胞影响许多生物学过程，包括胚胎发育、组织修复和再生、肿瘤抑制和衰老。衰老细胞的标志包括但不限于衰老相关的β-半乳糖苷酶活性增加（SA-β-gal）; p16 INK 4A、p53和p21水平;更高水平的DNA损伤，包括γ-H2 AX;衰老相关异染色质灶（SAHF）的形成;以及获得衰老相关分泌表型（SASP），一种以分泌大量促炎细胞因子和信号分子为特征的现象。在这里，我们描述了在培养细胞中复制和DNA损伤诱导衰老的协议。此外，我们强调了使用几种衰老相关标志物（包括SA-β-gal、γ-H2 AX和SAHF染色）监测衰老表型的技术，以及定量细胞周期调节因子和SASP因子的蛋白质和mRNA水平的技术。这些方法可应用于各种模型和组织的衰老评估。
In response to cellular stress or damage, proliferating cells can induce a specific program that initiates a state of long-term cell-cycle arrest, termed cellular senescence. Accumulation of senescent cells occurs with organismal aging and through continual culturing in vitro. Senescent cells influence many biological processes, including embryonic development, tissue repair and regeneration, tumor suppression, and aging. Hallmarks of senescent cells include, but are not limited to, increased senescence-associated β-galactosidase activity (SA-β-gal); p16INK4A, p53, and p21 levels; higher levels of DNA damage, including γ-H2AX; the formation of Senescence-associated Heterochromatin Foci (SAHF); and the acquisition of a Senescence-associated Secretory Phenotype (SASP), a phenomenon characterized by the secretion of a number of pro-inflammatory cytokines and signaling molecules. Here, we describe protocols for both replicative and DNA damage-induced senescence in cultured cells. In addition, we highlight techniques to monitor the senescent phenotype using several senescence-associated markers, including SA-β-gal, γ-H2AX and SAHF staining, and to quantify protein and mRNA levels of cell cycle regulators and SASP factors. These methods can be applied to the assessment of senescence in various models and tissues.
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影响因子：
64.8
作者：
Baker DJ;Childs BG;Durik M;Wijers ME;Sieben CJ;Zhong J;Saltness RA;Jeganathan KB;Verzosa GC;Pezeshki A;Khazaie K;Miller JD;van Deursen JM
通讯作者：
van Deursen JM
DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
癌基因诱导的DNA损伤反应与衰老和癌症异染色质之间的相互作用。
影响因子：
21.3
作者：
Di Micco R;Sulli G;Dobreva M;Liontos M;Botrugno OA;Gargiulo G;dal Zuffo R;Matti V;d'Ario G;Montani E;Mercurio C;Hahn WC;Gorgoulis V;Minucci S;d'Adda di Fagagna F
通讯作者：
d'Adda di Fagagna F
影响因子：
11.8
作者：
Demaria, Marco;Ohtani, Naoko;Youssef, Sameh A.;Rodier, Francis;Toussaint, Wendy;Mitchell, James R.;Laberge, Remi-Martin;Vijg, Jan;Van Steeg, Harry;Dolle, Martijn E. T.;Hoeijmakers, Jan H. J.;de Bruin, Alain;Hara, Eiji;Campisi, Judith
通讯作者：
Campisi, Judith
影响因子：
64.5
作者：
Narita, M;Nunez, S;Lowe, SW
通讯作者：
Lowe, SW