Method To Purify and Analyze Heterogeneous Senescent Cell Populations Using a Microfluidic Filter with Uniform Fluidic Profile

Kim, Minseok S.; Jo, Seong-Hyeon; Park, Taezoon; Kim, Sun Soo; Gurel, Ogan; Park, Sang Chul

doi:10.1021/acs.analchem.5b00445

Cited by 7 publications

(5 citation statements)

References 29 publications

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“…Plasma membranes, due to altered lipid composition, become more rigid with age (Fulop et al, 2012a;Momchilova et al, 2014). Moreover, senescent cells increase in size compared to non-senescent analogues (Kim et al, 2015;Rodier and Campisi, 2011). This modifies many essential functions within the cell, such as diffusion, cell size, membrane fusion, chemical and electrical processes, cell elasticity and membrane stiffness (Pontes et al, 2013).…”

Section: Plasma Membrane Lipid Compositionmentioning

confidence: 99%

Biomarkers to identify and isolate senescent cells

Matjusaitis

Chin

Sarnoski

et al. 2016

Ageing Research Reviews

123

108

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Highlights There is no single biomarker which can robustly identify senescent cells. Widely used senescent cell biomarkers should be used in combination for accuracy. Technologies like tangential flow filtration can be used to isolate senescent cells. Other methods, like senolytic viruses, could be used to remove senescent cells. AbstractAging is the main risk factor for many degenerative diseases and declining health. Senescent To better understand cell aging and to reap the benefits of senescent cell removal, it is necessary to have a reliable biomarker to identify these cells. Following an introduction to cellular senescence, we discuss several classes of biomarkers in the context of their utility in identifying and/or removing senescent cells from tissues. Although senescence can be induced by a variety of stimuli, senescent cells share some characteristics that enable their identification both in vitro and in vivo. Nevertheless, it may prove difficult to identify a single biomarker capable of distinguishing senescence in all cell types. Therefore, this will not be a comprehensive review of all senescence biomarkers but rather an outlook on technologies and markers that are most suitable to identify and isolate senescent cells.

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Section: Plasma Membrane Lipid Compositionmentioning

confidence: 99%

Biomarkers to identify and isolate senescent cells

Matjusaitis

Chin

Sarnoski

et al. 2016

Ageing Research Reviews

123

108

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“…However, few studies have considered the uniformity of flow velocity in CTC isolation technologies. The chip was specially designed to have a uniform flow along the length of the microslit based on our previous work [40]. To accomplish this, a diamond shape barrier was placed between the input and the slit (S1 Movie).…”

Section: Resultsmentioning

confidence: 99%

Microslit on a chip: A simplified filter to capture circulating tumor cells enlarged with microbeads

et al. 2019

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Microchips are widely used to separate circulating tumor cells (CTCs) from whole blood by virtues of sophisticated manipulation for microparticles. Here, we present a chip with an 8 μm high and 27.9 mm wide slit to capture cancer cells bound to 3 μm beads. Apart from a higher purity and recovery rate, the slit design allows for simplified fabrication, easy cell imaging, less clogging, lower chamber pressure and, therefore, higher throughput. The beads were conjugated with anti-epithelial cell adhesion molecules (anti-EpCAM) to selectively bind to breast cancer cells (MCF-7) used to spike the whole blood. The diameter of the cell-bead construct was in average 23.1 μm, making them separable from other cells in the blood. As a result, the cancer cells were separated from 5 mL of whole blood with a purity of 52.0% and a recovery rate of 91.1%, and also we confirmed that the device can be applicable to clinical samples of human breast cancer patients. The simple design with microslit, by eliminating any high-aspect ratio features, is expected to reduce possible defects on the chip and, therefore, more suitable for mass production without false separation outputs.

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“…30 We evaluated cell size, because membrane lipid composition and membrane biophysical properties are altered by cellular senescence, resulting in an alteration of cell size. 33,34 In addition, cyclin dependent kinase inhibitors (CKIs), such as p16 and p21, have been used as biomarkers, because cell cycle arrest is recognized as a hallmark of cellular senescence. 30 Indeed, overexpression of p21 promotes cellular senescence, 35 and downregulation of FIGURE 2.…”

Section: Discussionmentioning

confidence: 99%

The Effect of a p38 Mitogen-Activated Protein Kinase Inhibitor on Cellular Senescence of Cultivated Human Corneal Endothelial Cells

Hongo

Okumura

Nakahara

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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Citation: Hongo A, Okumura N, Nakahara M, Kay EP, Koizumi N. The effect of a p38 mitogen-activated protein kinase inhibitor on cellular senescence of cultivated human corneal endothelial cells. Invest Ophthalmol Vis Sci. 2017;58:3325-3334. DOI:10.1167/iovs.16-21170 PURPOSE. We have begun a clinical trial of a cell-based therapy for corneal endothelial dysfunction in Japan. The purpose of this study was to investigate the usefulness of a p38 MAPK inhibitor for prevention cellular senescence in cultivated human corneal endothelial cells (HCECs).METHODS. HCECs of 10 donor corneas were divided and cultured with or without SB203580 (a p38 MAPK inhibitor). Cell density and morphology were evaluated by phase-contrast microscopy. Expression of function-related proteins was examined by immunofluorescent staining. Cellular senescence was evaluated by SA-b-gal staining and Western blotting for p16 and p21. Senescence-associated factors were evaluated by membrane blotting array, quantitative PCR, and ELISA.RESULTS. Phase-contrast microscopy showed a significantly higher cell density for HCECs cultured with SB203580 than without SB203580 (2623 6 657 cells/mm 2 and 1752 6 628 cells/mm 2 , respectively). The HCECs cultured with SB203580 maintained a hexagonal morphology and expressed ZO-1, N-cadherin, and Na þ /K þ -ATPase in the plasma membrane, whereas the control HCECs showed an altered staining pattern for these marker proteins. HCECs cultured without SB203580 showed high positive SA-b-gal staining, a low nuclear/ cytoplasm ratio, and expression of p16 and p21. IL-6, IL-8, CCL2, and CXCL1 were observed at high levels in low cell density HCECs cultured without SB203580.CONCLUSIONS. Activation of p38 MAPK signaling due to culture stress might be a causative factor that induces cellular senescence; therefore, the use of p38 MAPK inhibitor to counteract senescence may achieve sufficient numbers of HCECs for tissue engineering therapy for corneal endothelial dysfunction.

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Method To Purify and Analyze Heterogeneous Senescent Cell Populations Using a Microfluidic Filter with Uniform Fluidic Profile

Cited by 7 publications

References 29 publications

Biomarkers to identify and isolate senescent cells

Biomarkers to identify and isolate senescent cells

Microslit on a chip: A simplified filter to capture circulating tumor cells enlarged with microbeads

The Effect of a p38 Mitogen-Activated Protein Kinase Inhibitor on Cellular Senescence of Cultivated Human Corneal Endothelial Cells

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