Differential responsiveness of early- and late-passage endothelial cells to shear stress

Kudo, Fabio A.; Warycha, B.; Juran, Peter J.; Asada, Hidenori; Teso, Desarom; Aziz, Faisal; Frattini, Jared C.; Sumpio, Bauer E.; Nishibe, Toshiya; Cha, Charles; Dardik, Alan

doi:10.1016/j.amjsurg.2005.07.017

Cited by 21 publications

(10 citation statements)

References 41 publications

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“…Enhanced apoptosis of porcine pulmonary artery endothelial cells (Zhang et al 2002) and declined arachidonic acid content and eicosanoid release in porcine aortic endothelial cells in response to agonist stimulation (Brown and Deykin 1991) were determined as well. The effects of cell passaging on endothelin-1 and prostaglandin F2a production of bovine luteal endothelial cells (Acosta et al 2007) and on the responsiveness of bovine aortic endothelial cells to orbital shear stress (Kudo et al 2005) were also evaluated. It has been reported that in vitro passaging causes the changes in enzyme activities and total protein in primate brain microvessel endothelial cells (Shi and Audus 1994) and in LDL uptake and expression of eNOS, MCP-1, von Willebrand factor (vWF), VCAM-1, ICAM-1, and E-selectin in primate femoral artery endothelial cells with or without cytokine stimulation (Shi et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells

Liao

Chen

et al. 2013

Cytotechnology

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The effects of serial cell passaging on cell spreading, migration, and cell-surface ultrastructures have been less investigated directly. This study evaluated the effects of long-term serial cell passaging (totally 35 passages) on cultured human umbilical vein endothelial cells which were pre-stored at -80°C as usual. Percentage-and spread area-based spreading assays, measurements of fluorescently labeled actin filaments, migration assay, and measurements of cellsurface roughness were performed and quantitatively analyzed by confocal microscopy or atomic force microscopy. We found that the abilities of cell spreading and migration first increased at early passages and then decreased after passage 15, in agreement with the changes in average length of actin filaments. Recovery from cold storage and effects of cell passaging were potentially responsible for the increases and decreases of the values, respectively. In contrast, the average roughness of cell surfaces (particularly the nucleussurrounding region) first dropped at early passages and then rose after passage 15, which might be caused by cold storage-and cell passaging-induced endothelial microparticles. Our data will provide important information for understanding serial cell passaging and implies that for pre-stored adherent cells at -80°C cell passages 5-10 are optimal for in vitro studies.

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Section: Introductionmentioning

confidence: 99%

Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells

Liao

Chen

et al. 2013

Cytotechnology

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“…Endothelial cells undergo senescence both in vivo and in vitro [1,2,3,4,5,6,7,8]. Since endothelial cells are a critical component of the vasculature, their senescence can have a significant impact on vascular integrity and function and on the progression of vascular disease [4,5].…”

Section: Introductionmentioning

confidence: 99%

Senescence of Cultured Porcine Coronary Arterial Endothelial Cells Is Associated with Accelerated Oxidative Stress and Activation of NFĸB

Lee

Wang

Vanhoutte³

2009

J Vasc Res

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Aims: Endothelial dysfunction occurs following multiple passaging in vitro,but the molecular mechanisms involved remain unidentified. The present study defined the genomic changes related to dysfunction in cultured senescent endothelial cells. Methods and Results: Senescent cells were produced by multiple passaging of porcine coronary arterial endothelial cells for up to 4 weeks. Genomic and proteomic studies on cultured cells at the first passage (P1) and the fourth passage (P4) were performed. Senescence and decreased NO production were observed in cells and several signaling pathways – such as IFN/STAT, IGF, TGF-β, cytoskeleton rearrangement and lipid metabolism – were altered at P4, as judged from the microarray analysis. The basal and stimulated (by TNF-α) levels of NFĸB were augmented in senescent cells in electrophoretic mobility shift assays in association with increased oxidative stress, increased p53 protein stability, and activated apoptotic pathways. The increased oxidative stress was alleviated by treatment with the superoxide dismutase mimetic MnTMPyP. Conclusions: After multiple passaging in vitro, porcine coronary endothelial cells exhibited dysfunction and senescence associated with reduced proliferative capacity, increased oxidative stress, and activation of the NFĸB and p53 signaling pathways.

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“…In addition, CD-selected hMSC after extended growth in culture showed a decline in shear response as has been observed for adherent selected cells at an earlier stage of culture/passage. 24 These variations demonstrate the inherent variability in using mixed populations of bone marrow–derived cells with quite marked cell responses to environmental cues; a feature which must be considered when designing cell therapies and sources of cells for use in therapeutic treatments.…”

Section: Discussionmentioning

confidence: 99%

Gene expression of single human mesenchymal stem cell in response to fluid shear

et al. 2012

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Stem cell therapy may rely on delivery and homing through the vascular system to reach the target tissue. An optical tweezer model has been employed to exert different levels of shear stress on a single non-adherent human bone marrow–derived mesenchymal stem cell to simulate physiological flow conditions. A single-cell quantitative polymerase chain reaction analysis showed that collagen type 1, alpha 2 (COL1A2), heat shock 70-kDa protein 1A (HSPA1A) and osteopontin (OPN) are expressed to a detectable level in most of the cells. After exposure to varying levels of shear stress, there were significant variations in gene transcription levels across human mesenchymal stem cells derived from four individual donors. Significant trend towards upregulation of COL1A2 and OPN gene expression following shear was observed in some donors with corresponding variations in HSPA1A gene expression. The results indicate that shear stress associated with vascular flow may have the potential to significantly direct non-adherent stem cell expression towards osteogenic phenotypic expression. However, our results demonstrate that these results are influenced by the selection process and donor variability.

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Differential responsiveness of early- and late-passage endothelial cells to shear stress

Cited by 21 publications

References 41 publications

Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells

Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells

Senescence of Cultured Porcine Coronary Arterial Endothelial Cells Is Associated with Accelerated Oxidative Stress and Activation of NFĸB

Gene expression of single human mesenchymal stem cell in response to fluid shear

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