“Senescent Cell Detection – Selection Guide”

Various Senescent Cell from Various Indicators

Product	Cellular Senescence Detection Kit - SPiDER-ßGal	Cellular Senescence Plate Assay Kit - SPiDER-ßGal	DNA Damage Detection Kit – γH2AX * Coming Soon	Nucleolus Bright Green / Red
Detection	Fluorescence	Fluorescence	Fluorescence	Fluorescence
Wavelength (Ex/Em)	500 - 540 nm / 530 - 570 nm	535 nm / 580 nm	Green: 494 nm / 518 nm Red: 550 nm / 566 nm Deep Red: 646 nm / 668 nm	Green: 513 nm / 538 nm Red: 537 nm / 605 nm
Indicator	SA-ß-gal activity	SA-ß-gal activity	Changes in DNA damage	Changes in the Nucleolus
Detection method	Imaging Substrate:SPiDER-ßGal	Plate assay Substrate:SPiDER-ßGal	Imaging Detection of γH2AX by secondary antibody method	Imaging Detection of the Nucleolus by RNA staining reagent
Instrument	Fluorescence microscopy Flow cytometry	Plate reader	Fluorescence microscopy	Fluorescence microscopy
Sample	Live cells Fixed cells	Live cells (lysis of live cells)	Fixed cells	Fixed cells
data
Item#	SG03	SG05	Green: G265 Red: G266 Deep Red: G267	Green: N511 Red: N512

Feature of Cellular Senescence

Cellular senescence is considered crucial to many different research areas, especially since the recent discovery of the senescence-associated secretory phenotype (SASP). SASP is a known risk factor for malignant transformation and exploration into stem cell research has found a link between SASP and the aging phenomenon. The features of cellular senescence were summarized based on modern publications that have a high number of citations.

Evaluate senescent cells from various makers

In WI-38 cells at different passages SA-ß-Gal activity, mitochondrial membrane potential, and cellular metabolism (Glucose and Lactate) were evaluated using each specific kit. In senescent cells, SA-ß-Gal activity was enhanced and mitochondrial membrane potential was reduced. Consumption of Glucose and lactate level in the supernatant measured as an indicator of metabolism was increased.

Sample	Senescence induction	Senescence Marker	Factor involved in senescence	Citation
IMR90 (human lung fibroblast cell)	passage of aging	SA-ßGal p16, p21 enlarged nucleoli	SETD8 expression ↓ H4K20me1 ↓ mitochondrial oxidative phosphorylation ↑ ribosome biogenesis ↑	①
	SETD8 (methyltransferase) low expression		mitochondrial oxidative phosphorylation ↑ ribosome biogenesis ↑
Aged satellite cells	-	SA-ßGal p16	Autophagy activity ↓ ROS ↑ Mitochondria membrane potential ↓	②
Atg7-deficient satellite cells	Inhibition of autophagy	SA-ßGal P15, p16, p21 γ-H2AX	ROS ↑ Mitochondria membrane potential ↓
Fibroblasts from patients with type 2 diabetes	-	SA-ßGal p21, p53 γ-H2AX	NADPH / NADP ↓ (resistance to oxidative stress ↓) NADPH oxidase ↑ (ROS ↑)	③
IMR90	Ethidium Bromide (depletion of mtDNA) + pyruvate withdrawal	SA-ßGal	NAD+ / NADH ↓	④
MDA-MB-231 (human breast cancer cells)	Radiation-induced senescence + low expression of securin	SA-ßGal	Lactate ↑ LDH activity ↑ (glycolysis ↑)	⑤
MEF (mouse embryonic fibroblasts)	overexpression of cancer gene passage of aging overexpression of transcription factor(E2F1)	SA-ßGal p16, p21 enlarged nucleoli	Ribosome RNA ↑ p53 ↑	⑥
Mouse tail-tip fibroblasts	2 month-old WT, 22 month-old WT, p16 KO(22 month-old)	SA-ßGal p14, p16	NAD+ ↓ SIRT3 ↓	⑦

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.