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Correction pp undefined-undefined
Correction for: Iron retardation in lysosomes protects senescent cells from ferroptosis
Relevance score: 9.279008Yujing Feng, Huaiqing Wei, Meng Lyu, Zhiyuan Yu, Jia Chen, Xinxing Lyu, Fengfeng Zhuang
Keywords: iron accumulation, senescent cells, lysosome, ferroptosis, ferritinophagy
Published in Aging on July 31, 2024
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Research Paper Volume 12, Issue 14 pp 15050-15057
Establishing a density-based method to separate proliferating and senescent cells from bone marrow stromal cells
Relevance score: 7.054573Fei Xu, Qiang Zhang, Haitao Wang
Keywords: bone marrow stromal cells, cell transplantation, density-based method, proliferating cells, senescent cells
Published in Aging on July 25, 2020
Isolation and Identification of BMSCs by Phenotypic Characterization and Multipotent Differentiation Potential. (A) Cells were isolated from the femurs and tibias of 3- to 4-week-old mice shown at P0 and P3. Cells are attached at P3. Scale Bar=200μm. (B) Flow cytometric analysis of cell surface markers on isolated BMSCs indicates Scal-1+ CD29+ CD11b- CD45- CD105-. (C) Differentiation capacity of BMSCs: ALP staining of cells cultured in osteogenic induction medium for 7 days (upper-left image); alizarin red staining of cells cultured in osteogenic induction medium for 21 days (upper-right image); oil red O staining of cells cultured in adipogenic induction medium for 7 days (lower-left image); and alcian blue staining of cells cultured in chondrogenic induction medium for 14 days (lower-right image). P0, passage 0; P2, passage 2; P3, passage 3. Scale Bar=200μm.
Density Gradient Separation of proliferating and Senescent Cells. (A) Immunofluorescence of BrdU positive cells is shown in the first panel. The right panel shows that the number of BrdU positive cells is significantly lower in P8 compared to P3. Scale Bar=200μm. (B) BMSCs (P8), a mixture of proliferating and senescent cells, were centrifuged through a density gradient medium (OptiPrep, Sigma-Aldrich) at 800g for 20 minutes. Aliquots (0.5 mL) were taken from the low- and high-density layers. The cells were then incubated in a 48-well plate. (C) SA-β-gal staining of the 2 groups. Low-density cells contained a higher percentage of SA-β-gal positive cells compared to the high-density cells. The right panel shows that the number of SA-β-gal positive cells is significantly lower in high-density cells compared to low-density cells (n=3). BrdU indicates bromodeoxyuridine; DAPI, 4′,6-diamidino-2-phenylindole; P3, passage 3; P8, passage 8; SA-β-gal, senescence-associated β-galactosidase; Paired T-Test **, P<.01. data are represented as mean ± SEM. Scale Bar=200μm.
Differentiation Capacity and TIF Assay of BMSCs After Separation. (A) Co-localization of 53BP1 DNA damage protein (green) and telomeric DNA (red). TIFs are indicated by arrows. Blue, DAPI; Red, Telomere; Orange/Yellow, TIFs. Scale Bar=5μm. (B) The percentage of TIF-positive cells is substantially lower in high-density cells compared to low-density cells (left panel). The protein levels of γH2AX and P21 were induced at low density (LD) using Western blot. P16 gene expression was increased at LD using real-time PCR. (C) Low- and high-density cells were cultured either in osteogenic induction medium or adipogenic induction medium for 7 days. ALP and oil red O staining were conducted at the end of 14 days. ALP staining (upper- and lower-left panels) was induced in the high-density cell group. Oil red O staining (upper- and lower-middle panels) was induced in the low-density cell group. Alizarin red staining (upper- and lower-right panels) was induced in the high-density cell group. Quantification of clones of oil red O staining positive (n=3) is shown in the graph at right. ALP indicates alkaline phosphatase; DAPI, 4′,6-diamidino-2-phenylindole; HD, high density; LD, low density; TIF, telomere dysfunction-induced foci. Paired T-Test, ***, P<.001, data are represented as mean ± SEM. Scale Bar=200μm.
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Research Paper Volume 8, Issue 11 pp 2915-2926
Discovery of piperlongumine as a potential novel lead for the development of senolytic agents
Relevance score: 7.7223887Yingying Wang, Jianhui Chang, Xingui Liu, Xuan Zhang, Suping Zhang, Xin Zhang, Daohong Zhou, Guangrong Zheng
Keywords: piperlongumine, aging, senescent cells, senolytic agents, ABT-263, reactive oxygen species, synergistic effect
Published in Aging on November 19, 2016
Senolytic activity of piperlongumine (PL). (A) Structures of PL, 2,3-dihydro-PL, and 7,8-dihydro-PL. (B) Quantification of viable WI-38 non-senescent cells (NC), IR-induced senescent cells (IR-SC), replication-exhausted senescent cells (Rep-SC), or Ras-induced senescent cells (Ras-SC) 72 h after treatment with increasing concentrations of PL (n = 3). (C) Quantification of viable IR-SCs over time after treatment with 10 µM PL (left) or after incubation with 10 µM PL, removal of the drug, and further culture for 72 h (right) (n = 3). (D) Quantification of viable WI-38 NCs and IR-SCs 72 h after treatment with increasing concentrations of 2,3-dihydro-PL or 7,8-dihydro-PL (n = 3). Data are represented as the mean ± SEM.
PL kills SCs by apoptosis. (A) Representative flow cytometric plots to measure apoptotic WI-38 IR-SCs at 48 h after treatment with vehicle (Veh), 10 µM PL, 10 µM Q-VD-Oph (QVD), or the combination of PL and QVD. (B) Quantification of the percentage of viable (gate II: PI− Annexin V−) and apoptotic (gates III and IV: PI− Annexin V+ and PI+ Annexin V+) (right) IR-SCs 48 h after treatment as in (A) (left), and quantification of the percentage of viable IR-SCs 72 h after treatment as in (A) (right). (C) Representative western blot and quantitative analysis of cleaved-poly(ADP-ribose) polymerase (cPARP), procaspase-3 (Procasp-3), cleaved caspase-3 (cCasp-3), and β-actin in NCs and WI-38 IR-SCs 24 h and 48 h after incubation with Veh or 10 µM PL. (D) Representative western blot analysis of RIP1, RIP3, and β-actin in WI-38 NCs and IR-SCs 24 h after incubation with Veh or 10 µM PL. A cell lysate of etoposide-treated Jurkat cells was used as a positive control. Data are represented as the mean ± SEM.
Effect of PL and its analogs on ROS production and senolytic activity in WI-38 IR-SCs. (A) Representative flow cytometric analysis of ROS production in NCs and IR-SCs 24 h after incubation with or without PL by DHR (left) (MFI, mean fluorescence intensity) and quantification of the fold increase of ROS levels in WI-38 NCs and WI-38 IR-SCs cells at the indicated times (middle and right) after incubation with 10 µM PL. As a positive control, cells were treated with 100 µM of H2O2 for 2 h, the H2O2 was removed, and cells were cultured for an additional 24 h (n = 3). (B) Quantification of the fold increase in DHR-123 MFI (left) in WI-38 IR-SCs 24 h after treatment with Veh, 10 µM PL, 2 mM NAC (pretreatment overnight), or the combination of PL and NAC, and (right) the percentage of viable WI-38 IR-SCs 72 h after treatment with Veh, 10 µM PL, 2 mM NAC (pretreatment overnight), or the combination of PL and NAC (n = 3). (C) Structure of PL-NAC and (Left) quantification of viable WI-38 NCs and WI-38 IR-SCs 72 h after treatment with increasing concentrations of PL-NAC (n = 3). (Right) Percentage of 10 µm PL remaining in the culture medium vs. time with or without 2mM NAC. (D) Left panel: quantification of the fold increase in DHR MFI (left) of WI-38 IR-SCs 24 h after treatment with Veh, 10 µM PL, 5 µM γ-tocotrienol (GT3, pretreatment overnight), or the combination of PL and GT3; and right panel: the percentage of viable WI-38 IR-SCs 72 h after treatment with Veh, 10 µM PL, 5 µM GT3 (pretreatment overnight), or the combination of PL and GT3 (n = 3). (E-H) Quantification of the fold increase in DHR-123 MFI after 24 h treatment (left) and viability of WI-38 NCs and WI-38 IR-SCs 72 h treatment (right) after they were treated with increasing concentrations or (E)10 µM BRD4809, (F) 0.5 µM PL-DI, (G) 0.625 µM PL-FPh, and (H) 5 µM PL-7 (n = 3). Data are represented as the mean ± SEM.
PL synergistically and selectively kills SCs in combination with ABT-263. (A) Quantification of NC viability 72 h after the cells incubation with vehicle, 1.25 µM ABT-263, 10 µM PL, or the combination of ABT-263 and PL (n = 3). (B) Quantification of WI-38 IR-SC viability 72 h after incubation with vehicle, 1.25 µM ABT-263, 5 or 10 µM PL, or the combination of ABT-263 and PL (n = 3-5). (C) Quantification of WI-38 IR-SC viability 72 h after incubation with vehicle, 10 µM PL, 0.08-1.25 µM ABT-263, or the combination of ABT-263 and PL (n = 3-6). Data are represented as the mean ± SEM.
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Research Perspective Volume 3, Issue 2 pp 168-174
Recent developments in the use of γ -H2AX as a quantitative DNA double-strand break biomarker
Relevance score: 7.1498814Christophe E. Redon, Asako J. Nakamura, Olga A. Martin, Palak R. Parekh, Urbain S. Weyemi, William M. Bonner
Keywords: DNA damage, DNA repair, γ -H2AX, double-strand break, biomarker, cancer, senescent cells
Published in Aging on February 11, 2011
Because of its sensitivity, the γ-H2AX assay is now utilized in many research areas “from benchtop to bedside” by researchers and clinicians. In addition to being widely used for fundamental research (study of genome stability, DNA repair, etc.) in the last decade, γ-H2AX was identified as a biomarker for cancer (and premalignant lesions) and used to better understand aging. Additionally, γ-H2AX has been developed for radiation biology and biodosimetry for drug development and clinical studies (chemotherapy, the impact of chronic inflammation and diabetes on genome integrity). Finally, γ-H2AX measurement is an efficient and sensitive genotoxic assay for environmental studies.
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Research Paper pp undefined-undefined
Iron retardation in lysosome protects senescent cells from ferroptosis
Relevance score: 10.048427Yujing Feng, Huaiqing Wei, Meng Lyu, Zhiyuan Yu, Jia Chen, Xinxing Lyu, Fengfeng Zhuang
Keywords: iron accumulation, senescent cells, lysosome, ferroptosis, ferritinophagy
Published in Aging on Invalid Date
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Review pp undefined-undefined
Senescent cell-derived vaccines: a new concept towards an immune response against cancer and aging?
Relevance score: 8.441129João Pessoa, Sandrina Nóbrega-Pereira, Bruno Bernardes de Jesus
Keywords: cancer, immunotherapy, tumor-associated senescent cells, senescence, antigen, vaccine
Published in Aging on Invalid Date