Endothelial characteristics in healthy endothelial colony forming cells; generating a robust and valid ex vivo model for vascular disease

Boer, Suzan de; Bowman, Mackenzie; Notley, Colleen; Mo, Aomei; Lima, Patrícia; Jong, Annika de; Dirven, Richard; Weijers, Ester M.; Lillicrap, David; James, Paula D.; Eikenboom, Jeroen

doi:10.1111/jth.14998

Cited by 19 publications

(32 citation statements)

References 37 publications

(57 reference statements)

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“…As a result of the impregnation with mango leaf extract, the number of cells on the surface of the disks increased notably, and so did the quality of the culture. Figure 8b,c impregnated at 35 • C and 100 bar and 400 bar, respectively, show that the endothelial cells present a more elongated and polygonal morphology, which is characteristic of healthy endothelial cells [28]. This cell morphology and distribution suggests that the coating of the polymers with the extract provides optimal conditions for the growth and maintenance of the culture.…”

Section: Cell Viabilitymentioning

confidence: 89%

Supercritical Impregnation of Mango Leaf Extract into PLA 3D-Printed Devices and Evaluation of Their Biocompatibility with Endothelial Cell Cultures

et al. 2022

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The addition of natural substances with pharmacoactive properties to polymeric biomedical devices would provide beneficial regarding the assimilation of these endoprostheses when implanted into a patient’s body. The added drug would facilitate endothelization by regulating the inflammatory processes that such interventions entail, preventing contamination hazards and favoring the angiogenesis or formation of blood vessels in the tissue. The present work used mango leaf extract (MLE) obtained through pressurized ethanol for this purpose. Polylactic acid (PLA) in the form of filaments or 3D-printed disks was impregnated by means of supercritical technology with MLE for the culture essays. The release kinetics has been studied and the polymer matrices have been examined by scanning electron microscopy (SEM). The impregnated devices were subjected to in vitro culture of colony-forming endothelial cells. The influence of the different impregnation conditions used for the production of the MLE impregnated polymeric devices on the development of the cell culture was determined by fluorescence microscopy. The best results were obtained from the calcein cultures on 35 °C MLE impregnated into 3D-printed polymer disks.

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Section: Cell Viabilitymentioning

confidence: 89%

Supercritical Impregnation of Mango Leaf Extract into PLA 3D-Printed Devices and Evaluation of Their Biocompatibility with Endothelial Cell Cultures

et al. 2022

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“…First, since the initial ECFC isolations and subsequent experiments, new reports have been published highlighting the variability and clonal properties of ECFCs. 21,22 It is possible that some of the variability observed in this study within families and/or the discrepancies between clinical phenotype and cellular phenotype may be a result of variations in clones and also not having taken into account the comparison of cells of the same morphological type. 22 The discrepancy between basal secretion from ECFCs and plasma VWF levels of patients (i.e., II-3) may also be attributed to faster clearance in the patient, which is not recapitulated in the cellular model.…”

Section: Discussionmentioning

confidence: 99%

“…21,22 It is possible that some of the variability observed in this study within families and/or the discrepancies between clinical phenotype and cellular phenotype may be a result of variations in clones and also not having taken into account the comparison of cells of the same morphological type. 22 The discrepancy between basal secretion from ECFCs and plasma VWF levels of patients (i.e., II-3) may also be attributed to faster clearance in the patient, which is not recapitulated in the cellular model. 11 Additionally, while VWF secretion from ECFCs has been shown to remain stable up to passage 10, 11 we do acknowledge that variability may have been due to differences in passage number of cells.…”

Section: Discussionmentioning

confidence: 99%

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von Willebrand factor propeptide variants lead to impaired storage and ER retention in patient‐derived endothelial colony‐forming cells

Bowman

Casey

Selvam

et al. 2022

Journal of Thrombosis and Haemostasis

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Background von Willebrand factor (VWF) is synthesized by vascular endothelial cells and megakaryocytes. The VWF propeptide is critical for multimerization and acts as an intra‐molecular chaperone for mature VWF in sorting to its storage organelles, Weibel‐Palade bodies (WPBs). In the Canadian Type 3 VWD study, almost half of the identified variants were in the VWF propeptide and these were associated with an increased bleeding phenotype. Objective To investigate VWF propeptide variants that cause quantitative von Willebrand disease (VWD) by utilizing patient‐derived endothelial colony‐forming cells (ECFCs). Patients/Methods Endothelial colony‐forming cells were isolated from five Type 3 VWD patients from four families with the following variants: (1) homozygous p.Asp75_Gly178del (deletion of exons 4 and 5 deletion; Ex4‐5del), (2) homozygous p.Cys633Arg, (3) homozygous p.Arg273Trp, and (4) p.Pro293Glnfs*164 and p.Gln419* inherited in the compound heterozygous state. Additionally, ECFCs were isolated from six family members (two Type 1 VWD, four unaffected). Results Endothelial colony‐forming cells from the Type 3 patient with the compound heterozygous genotype exhibited a true null VWF cellular phenotype, with negligible VWF detected. In contrast, the other three propeptide variants presented a similar expression pattern in homozygous ECFCs where VWF was synthesized but not packaged in WPBs, and variant VWF had an increased association with the endoplasmic reticulum (ER) marker, protein disulfide‐isomerase (PDI), indicating an ER‐retention phenotype. The biosynthetic phenotype was similar but to a lesser degree in heterozygous ECFCs expressing the non‐null variants. Conclusion This study further elucidates the importance of the VWF propeptide in the VWD phenotype using patient‐derived cells.

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“…In vitro cell models can narrow the gap between in vitro and in vivo study of vascular diseases [ 2 ]. These models have a major role in studying the influence of various factors on vascular diseases [ 3 ]. In addition, they are useful for studying drug–drug interaction, drug absorption, distribution, metabolism, excretion, and toxicity testing, or drug screening [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

A Potential In Vitro 3D Cell Model to Study Vascular Diseases by Simulating the Vascular Wall Microenvironment and Its Application

Deng

Hao

et al. 2022

Life

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Background: Current in vitro vascular models are too simple compared with the real vascular environment. In this research, a novel in vitro 3D vascular disease model that simulated the vascular microenvironment was introduced. Methods: This model was mainly established by low shear stress and co-culture of endothelial cells and smooth muscle cells. Characterization and reproduction of the pathological state of the 3D model were determined. The effect of two clinical drugs was verified in this model. The difference of drug screening between a traditional oxidative-damaged cell model and this 3D model was determined by HPLC. Results: This model presented many disease markers of vascular diseases: abnormal cellular shape, higher endothelial cell apoptotic rate and smooth muscle cell migration rate, decreased superoxide dismutase level, and increased malondialdehyde and platelet-derived growth factor level. The drugs effectively reduced the disease indices and relieved the damage caused by low shear stress. Compared to the traditional oxidative-damaged cell model, this 3D model screened different active components of Salviae Miltiorrhizae extract, and it is closer to clinical studies. Conclusions: These results suggest that the 3D vascular disease model is a more efficient and selective in vitro study and drug screening platform for vascular diseases than previously reported in vitro vascular disease models.

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Endothelial characteristics in healthy endothelial colony forming cells; generating a robust and valid ex vivo model for vascular disease

Abstract: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

Cited by 19 publications

References 37 publications

Supercritical Impregnation of Mango Leaf Extract into PLA 3D-Printed Devices and Evaluation of Their Biocompatibility with Endothelial Cell Cultures

Supercritical Impregnation of Mango Leaf Extract into PLA 3D-Printed Devices and Evaluation of Their Biocompatibility with Endothelial Cell Cultures

von Willebrand factor propeptide variants lead to impaired storage and ER retention in patient‐derived endothelial colony‐forming cells

A Potential In Vitro 3D Cell Model to Study Vascular Diseases by Simulating the Vascular Wall Microenvironment and Its Application

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