Cell Viability and Prostacyclin Release in Cultured Human Umbilical Vein Endothelial Cells

Rodríguez-Morata, A.; Garzón, Ingrid; Alaminos, Miguel; García‐Honduvilla, Natalio; Sánchez-Quevedo, M.C.; Buján, Julia; Campos, Antonìo

doi:10.1016/j.avsg.2008.03.004

Cited by 15 publications

(18 citation statements)

References 47 publications

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“…For these reasons, a deep study of sequential cell passages of cultured human TMJF might be a useful tool for tissue engineers in order to select the most suitable cell passage in terms of cell viability and differentiation from a clinical standpoint. In fact, several previous studies previously demonstrated that cell viability may vary among several cell passages and that selection of the most adequate cell passage is very important for cell therapy success [10], [11].…”

Section: Introductionmentioning

confidence: 99%

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

et al. 2012

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Temporo-mandibular joint disc disorders are highly prevalent in adult populations. Autologous chondrocyte implantation is a well-established method for the treatment of several chondral defects. However, very few studies have been carried out using human fibrous chondrocytes from the temporo-mandibular joint (TMJ). One of the main drawbacks associated to chondrocyte cell culture is the possibility that chondrocyte cells kept in culture tend to de-differentiate and to lose cell viability under in in-vitro conditions. In this work, we have isolated human temporo-mandibular joint fibrochondrocytes (TMJF) from human disc and we have used a highly-sensitive technique to determine cell viability, cell proliferation and gene expression of nine consecutive cell passages to determine the most appropriate cell passage for use in tissue engineering and future clinical use. Our results revealed that the most potentially viable and functional cell passages were P5–P6, in which an adequate equilibrium between cell viability and the capability to synthesize all major extracellular matrix components exists. The combined action of pro-apoptotic (TRAF5, PHLDA1) and anti-apoptotic genes (SON, HTT, FAIM2) may explain the differential cell viability levels that we found in this study. These results suggest that TMJF should be used at P5–P6 for cell therapy protocols.

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Section: Introductionmentioning

confidence: 99%

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

et al. 2012

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“…The principle reason for using these cells as surrogates for the respective natural cells is because of the variability of the cells from natural sources, [75][76][77][78][79][80][81] which in combination with the high variability of human donor blood would lead to uninterpretable data. The reproducibility problem for endothelial cells from various natural sources and from passage to passage is well documented.…”

Section: Discussionmentioning

confidence: 99%

Cellular regulation of blood coagulation: a model for venous stasis

Campbell

Brummel‐Ziedins

Butenas

et al. 2010

Blood

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We have adapted the corn-trypsin inhibitor whole-blood model to include EA.hy926 as an endothelium surrogate to evaluate the vascular modulation of blood coagulation initiated by relipidated recombinant tissue factor (rTf) and a cellular Tf surrogate, lipopolysaccharide (LPS)-stimulated THP1 cells (LPS-THP-1). Compared with bare tubes, EA.hy926 with rTf decreased the rate of thrombin formation, ITS accumulation, and the production of fibrinopeptide A. These phenomena occurred with increased rates of factor Va (fVa) inactivation by cleavages at R 506 and R 306 . Thus, EA.hy926 provides thrombin-dependent protein C activation and APC fVa inactivation. Comparisons of rTf with LPS-THP-1 showed that the latter gave reduced rates for TAT formation but equivalent fibrinopeptide A, and fV activation/inactivation. In the presence of EA.hy926, the reverse was obtained; with the surrogate endothelium and LPS-THP-1 the rates of TAT generation, fibrinopeptide release, and fV activation were almost doubled, whereas cleavage at R 306 was equivalent. These observations suggest cooperativity between the 2 cell surrogates. These data suggest that the use of these 2 cell lines provides a reproducible quasiendothelial quasi-inflammatory cytokinestimulated monocyte system that provides a method to evaluate the variations in blood phenotype against the background of stable inflammatory cell activator and a stable vascular endothelial surrogate.(Blood. 2010; 116(26):6082-6091) IntroductionVirchow postulated 3 phenomena that lead to venous thrombosis: (1) the components contained within the blood itself lead to systemic hypercoagulability; (2) alteration of the endothelium from injury or dysfunction; and (3) alterations of blood flow. 1 These postulates are the foundation for the physiologic understanding of blood coagulation and the maintenance of the hemostatic balance in the venous circulation.The maintenance of blood fluidity and vascular integrity is actively maintained by collaborations between the vascular wall and the procoagulant and anticoagulant functions in blood. Cellbound and soluble constituents cooperate to maintain this quasiequilibrium fluid state. Upon vascular perforation, multiple interactions initiate a local coagulant response, promoting platelet adhesion/ aggregation, the generation of membrane-bound catalysts, thrombin generation, and fibrin deposition that overcome local anticoagulants. These processes arrest blood loss with dimensions relevant to the injury. Subsequently, fibrinolysis and cellular proliferation repair 2 the injured tissue.The coagulation processes have been studied in vitro using anticoagulated phlebotomy blood, 3 derived plasma, 4,5 isolated component systems, [6][7][8] and numerical simulations. 9,10 We have used corn-trypsin inhibitor (CTI) stabilized whole blood to study the tissue factor (Tf)-initiated process in the absence of chelating anticoagulants. 11,12 Several hundred of these experiments have been performed to characterize the temporal dynamics of various blood clotting reactions...

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“…To view the film (PAH-PSS) 3 -PAH deposition into the arterial lumen, the polycation (PAH) of the last layer was coupled to fluorochrome fluorescence green: FITC (fluorescein iso cyanate Thio) and observed by confocal laser scanning microscopy (method described by Kerdjoudj and colleagues [2]). …”

Section: Characterization By Confocal Microscopymentioning

confidence: 99%

“…In this respect, autologous vessels remain the materials of choice for arterial reconstruction and bypass procedures because of the natural compliancy of the vessels and the antithrombogenic potential induced by endothelial cells. However, autografts can be unsuitable or unavailable [2,3], thus replacement proves to be problematic. The development of a functional smalldiameter vascular graft has long been a "holy grail".…”

Section: Introductionmentioning

confidence: 99%

“…The development of a functional smalldiameter vascular graft has long been a "holy grail". According to the tissue engineering paradigm, cells are seeded on a scaffold composed of synthetic polymer or natural material [2,3]. Then, the tissue is matured in vitro and the construct is implanted in the appropriate anatomic location as prosthesis [2].…”

Section: Introductionmentioning

confidence: 99%

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Brillouin spectroscopy: A new tool to decipher viscoelastic properties of biological scaffold functionalized with nanoscale films

Beroud

Vincent

Paternotte

et al. 2013

Bio-Medical Materials and Engineering

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BACKGROUND: In tissue engineering, the endothelialization of vascular scaffold can be a crucial step to improve graft patency. A functional cellularization requires coating surfaces. Since 2003, our group used polyelectrolyte multilayer films (PEMFs) made of poly(allylamine hydrochloride) and polystyren sulfonate to coat luminal surface of blood vessel. Previous results showed that PEMFs have remarkable effect on cellular behavior: adhesion, proliferation, differentiation. However, no method seems adapted for in vitro measurement of the viscoelastic shift after PEMFs buildup. OBJECTIVE: In this present work, we proposed to use a new analytical method based on Brillouin spectroscopy (BS) to investigate the influence PEMFs coating on vessel intrinsic viscoelasticy. METHODS: On human umbilical arteries and rabbit vessels, PEMFs were buildup and the luminal surfaces viscoelasticy were measuring by BS. RESULTS: It seems that these films do not alter dynamic functionality and BS could be an interesting method for understanding the role of the tissue architecture, the interrelation between the different structures constituting the wall and the influence of this architecture on the tissue behavior, especially with the characterized components of the different vascular wall. CONCLUSION: The ability of BS to characterize biological samples opens potential applications in tissue engineering field, especially as a tool for a better understanding of vascular diseases.

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Cell Viability and Prostacyclin Release in Cultured Human Umbilical Vein Endothelial Cells

Cited by 15 publications

References 47 publications

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

Cellular regulation of blood coagulation: a model for venous stasis

Brillouin spectroscopy: A new tool to decipher viscoelastic properties of biological scaffold functionalized with nanoscale films

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