Senescent Cells and Lysosomal Cholesterol Accumulation DOJINDO LABORATORIES

Previous Science Note

This article focusing on the role of cholesterol accumulation in lysosomes of senescent cells, and its connection to maintaining the senescence-associated secretory phenotype (SASP). We believe that these findings can provide valuable insights into the relationship between senescence and lysosomal function.
Lysosomal control of senescence and inflammation through cholesterol partitioning Kyeonghwan Roh, et. al., Nature Metabolism (2023) Point of Interest - Cholesterol is essential for various cellular functions, but its dysregulation can lead to age-related pathologies. - Senescent cells have been found to accumulate cholesterol in lysosomes, which helps maintain their SASP. - The accumulation of lysosomal cholesterol results in the formation of cholesterol-rich microdomains that sustain mTORC1 activity, supporting SASP. - By modulating lysosomal cholesterol partitioning, researchers were able to alter senescence-associated inflammation and osteoarthritis progression in mice, suggesting a potential regulatory role for cholesterol in age-related inflammation.
Related Techniques
Cellular senescence detection (Live cell imaging or FCM)	Cellular Senescence Detection Kit HOT
Cellular senescence detection (Plate reader)	Cellular Senescence Plate Assay Kit HOT
Autophagy detection	DAPGreen / DAPRed (Autophagosome detection), DALGreen (Autolysosome detection)
Lysosomal function assay	Lysosomal pH and mass detection Kit
Lysosome staining	pH-dependent (Red) and pH-independent (Green / Deep Red) probes
Nutrient uptake Assay	Glucose (Blue / Green / Red), Amino Acid, Cystine, and Fatty Acid Uptake Assay Kit
Lipid droplets detection	Lipi-Blue / Green / Red / Deep Red
Related Applications
Analysis of Lysosomal Mass and pH Exchange in Senescence-induced Cells
	Purpose: To investigate changes in lysosomal mass and pH in A549 cells induced to senescence by treatment with Doxorubicin (DOX). Methods: Senescence-associated acidic β-galactosidase (SA-βGal) activity was detected using Cellular Senescence Detection Kit - SPiDER-βGal. Lysosomal mass was detected using LysoPrime Deep Red, and pH was detected using pHLys Red. Fluorescence imaging was used to observe changes in lysosomal mass and pH in senescent cells compared to non-senescent cells. The normalized fluorescence intensity of lysosomal mass and pH was also measured by a plate reader. Results: Our findings indicate that senescence induced by DOX resulted in an increase in lysosomal mass and acidification of pH compared to non-senescent cells. The obtained results are consistent with previous reports* that demonstrated enhanced lysosomal activity in senescent cells induced by the CDK4/6 inhibitor, palbociclib. The fluorescence imaging and plate reader data both support these findings. * Miguel Rovira , et. al., Aging Cell (2022) ＜Experimental Conditions for Microscopy＞ SA-βGal(Green):Ex = 488 nm, Em = 490 – 550 nm Lysosomal pH (Red):Ex = 561 nm, Em = 560 – 620 nm Lysosomal mass (Deep Red):Ex = 633 nm, Em = 640 – 700 nm ＜Experimental Conditions for Plate Reader＞ SA-βGal: Ex = 525 – 535 nm, Em = 550 – 570 nm Lysosomal pH: Ex = 555 – 565 nm, Em = 590 – 610 nm Lysosomal mass: Ex = 645 – 655 nm, Em = 690 – 710 nm <Products in Use> - Cellular Senescence Detection Kit - Lysosomal pH and mass detection Kit More about Lysosomal Function Analysis