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2021/08/31

【Dojindo】細胞衰老指標選擇指南



細胞衰老(Cellular senescence)被認為對許多不同的研究領域至關重要，特別是自從近期發現衰老相關分泌蛋白(Senescence-associated secretory phenotype, SASP)以來。SASP是已知的惡性轉化風險因素，對幹細胞研究的探索發現SASP與衰老現象之間具有關聯性。

細胞衰老指標的偵測
Product Cellular Senescence Detection Kit
– SPiDER-βGal		 Cellular Senescence Plate Assay Kit - SPiDER-βGal DNA Damage
Detection Kit
–γH2AX		 Nucleolus Bright Green / Red
Cat. No. SG03 SG05 Green: G265 / Red:G266 / Deep Red: G267 Green: N511 / Red: N512
Wavelength
(Ex/Em)		 500 - 540 nm /
530 - 570 nm		 535 nm / 580 nm Green: 494 nm / 518 nm Green: 513 nm / 538 nm
Red: 550 nm / 566 nm Red: 537 nm / 605 nm
Deep Red: 646 nm / 668 nm -
指標 SA-β-gal activity SA-β-gal activity Changes in DNA Damage Changes in the Nucleolus
偵測法 Imaging Plate Assay Imaging Detection of 
γH2AX		 Imaging Detection of the
Substrate: SPiDER-βGal Substrate:
SPiDER-βGal		 secondary antibody method Nucleolus by RNA staining reagent
適用偵測平台 Fluorescence Microscopy Plate Reader Fluorescence Microscopy  Fluorescence Microscopy
Flow cytometry
適用偵測樣本 Live Cells, Fixed Cells Live cells (Lysis of live cells) Fixed cells Fixed cells
Data



細胞衰老的特徵




通過不同指標評估衰老細胞
在不同細胞代數的WI-38細胞中，使用不同的檢測試劑盒分別評估細胞的SA-ß-Gal活性，粒線體膜電位和細胞代謝情況（葡萄糖和乳酸）。在衰老細胞中，SA-ß-Gal活性增強，粒線體膜電位降低。細胞培養上清液中葡萄糖和乳酸的消耗量等代謝指標有所增加。





細胞老化導致相關因子的變化



Publications
① H. Tanaka, S. Takebayashi, A. Sakamoto, N. Saitoh, S. Hino and M. Nakao, “The SETD8/PR-Set7 Methyltransferase Functions as a Barrier to Prevent Senescence-Associated Metabolic Remodeling.”, Cell Reports, 2017, 18(9), 2148.
② L. Garcia-Prat, M. Martinez-Vicente and P. Munoz-Canoves, “Autophagy: a decisive process for stemness”, Oncotarget, 2016, 7(11), 12286.
③ M. Bitar, S. Abdel-Halim and F. Al-Mulla, “Caveolin-1/PTRF upregulation constitutes a mechanism for mediating p53-induced cellular senescence: implications for evidence-based therapy of delayed wound healing in diabetes”, Am J Physiol Endocrinol Metab., 2013, 305(8), E951.
④ C. Wiley, M. Velarde, P. Lecot, A. Gerencser, E. Verdin, J. Campisi, et. al., “Mitochondrial Dysfunction Induces Senescence with a Distinct Secretory Phenotype”, Cell Metab., 2016, 23(2), 303.
⑤ E. Liao, Y. Hsu, Q. Chuah, Y. Lee, J. Hu, T. Huang, P-M Yang & S-J Chiu, “Radiation induces senescence and a bystander effect through metabolic alterations.”, Cell Death Dis., 2014, 5, e1255.
⑥ K. Nishimura, T. Kumazawa, T. Kuroda, A. Murayama, J. Yanagisawa and K. Kimura, “Perturbation of Ribosome Biogenesis Drives Cells into Senescence through 5S RNP-Mediated p53 Activation”, Cell Rep. 2015, 10(8), 1310.
⑦ M. J. Son, Y. Kwon, T. Son and Y. S. Cho, “Restoration of Mitochondrial NAD+ Levels Delays Stem Cell Senescence and Facilitates Reprogramming of Aged Somatic Cells”, Stem Cells. 2016, 34(12), 2840.