A Novel Method for Isolating Pure Microvascular Endothelial Cells from Subcutaneous Fat Tissue Ideal for Direct Cell Seeding

Arts, C.H.; Heijnen-Snyder, G.J.; Joosten, P.Ph.A. Hedeman; Verhagen, Hence J.M.; Eikelboom, B.C.; Sixma, Jan J.; Groot, Philip G. de

doi:10.1038/labinvest.3780360

Cited by 13 publications

(4 citation statements)

References 6 publications

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“…This is similar to peripheral vein harvest approaches previously reported for isolation of ECs for vascular graft seeding (13,30). Autologous ECs could also be isolated more rapidly from adipose tissue (31) or circulating blood (32,33), which could reduce the patient's wait time for endothelialization from weeks to days or possibly even to hours.…”

Section: Discussionsupporting

confidence: 58%

Readily Available Tissue-Engineered Vascular Grafts

et al. 2011

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Autologous or synthetic vascular grafts are used routinely for providing access in hemodialysis or for arterial bypass in patients with cardiovascular disease. However, some patients either lack suitable autologous tissue or cannot receive synthetic grafts. Such patients could benefit from a vascular graft produced by tissue engineering. Here, we engineer vascular grafts using human allogeneic or canine smooth muscle cells grown on a tubular polyglycolic acid scaffold. Cellular material was removed with detergents to render the grafts nonimmunogenic. Mechanical properties of the human vascular grafts were similar to native human blood vessels, and the grafts could withstand long-term storage at 4 °C. Human engineered grafts were tested in a baboon model of arteriovenous access for hemodialysis. Canine grafts were tested in a dog model of peripheral and coronary artery bypass. Grafts demonstrated excellent patency and resisted dilatation, calcification, and intimal hyperplasia. Such tissue-engineered vascular grafts may provide a readily available option for patients without suitable autologous tissue or for those who are not candidates for synthetic grafts.

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Section: Discussionsupporting

confidence: 58%

Readily Available Tissue-Engineered Vascular Grafts

et al. 2011

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“…30–32 To examine the expression of miR-181b, an anti-inflammatory microRNA that we previously identified in the macrovasculature 33, 34 , ECs or adipocytes were isolated as described 21, 35–37 from eWAT of C57BL/6 mice fed a 60% HFD for 0, 3, 7, or 14 days followed by qPCR analysis (Figure 1A). We found miR-181b is the most dominantly expressed among miR-181 family members in adipose tissue ECs as demonstrated that its expression is 3.6-fold higher than that of miR-181a.…”

Section: Resultsmentioning

confidence: 99%

MicroRNA-181b Improves Glucose Homeostasis and Insulin Sensitivity by Regulating Endothelial Function in White Adipose Tissue

Sun

Lin

Zhang

et al. 2016

Circulation Research

118

100

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Rationale The pathogenesis of insulin resistance involves dysregulated gene expression and function in multiple cell types including endothelial cells (ECs). Posttranscriptional mechanisms such as microRNA-mediated regulation of gene expression could affect insulin action by modulating EC function. Objective To determine whether microRNA-181b (miR-181b) affects the pathogenesis of insulin resistance by regulating EC function in white adipose tissue during obesity. Methods and Results MiR-181b expression was reduced in adipose tissue ECs of obese mice, and rescue of miR-181b expression improved glucose homeostasis and insulin sensitivity. Systemic intravenous delivery of miR-181b robustly accumulated in adipose tissue ECs, enhanced insulin-mediated Akt phosphorylation at Ser473, and reduced endothelial dysfunction, an effect that shifted macrophage polarization towards an M2 anti-inflammatory phenotype in epididymal white adipose tissue (eWAT). These effects were associated with increased eNOS and FoxO1 phosphorylation as well as nitric oxide activity in eWAT. In contrast, miR-181b did not affect insulin-stimulated Akt phosphorylation in liver and skeletal muscle. Bioinformatics and gene profiling approaches revealed that PHLPP2, a phosphatase that dephosphorylates Akt at Ser473, is a novel target of miR-181b. Knockdown of PHLPP2 increased Akt phosphorylation at Ser473 in ECs, and ‘phenocopied’ miR-181b’s effects on glucose homeostasis, insulin sensitivity, and inflammation of eWAT in vivo. Finally, ECs from diabetic subjects exhibited increased PHLPP2 expression. Conclusions Our data underscore the importance of adipose tissue EC function in controlling the development of insulin resistance. Delivery of miR-181b or PHLPP2 inhibitors may represent a new therapeutic approach to ameliorate insulin resistance by improving adipose tissue endothelial Akt-eNOS-NO signaling.

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“…In previous work Arts and co‐workers [12] have used magnetic beads to remove both fibroblast and smooth muscle cells for cell purification. Our experience with CDw90 beads has shown that fibroblasts can be removed but the results are inconsistent and currently this is not a reliable technique for clinical use.…”

Section: Discussionmentioning

confidence: 99%

Extraction of cells for single‐stage seeding of vascular‐bypass grafts

Tiwari

Kidane

Punshon

et al. 2003

Biotech and App Biochem

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Experimental data are reported for the seeding of prosthetic vascular grafts with either mesothelial or endothelial cells as part of a research strategy in tissue engineering with the aim of improving graft patency and developing new techniques for single-stage cell extraction and seeding that would give a step reduction in surgery time. New data are reported for two different sources of cells, peritoneal lavage and subcutaneous fat. All experiments were undertaken in patients undergoing abdominal aortic aneurysm repair. Cells extracted from peritoneal lavage were insufficient for a single-stage seeding process. Subcutaneous fat was processed using either a positive cell-extraction method using CD31 Dynabeads or by a negative extraction method using CDw90-coated magnetic beads. Only positive cell extraction gave reliably sufficient numbers of endothelial cells as a source for single-stage seeding of vascular grafts.

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A Novel Method for Isolating Pure Microvascular Endothelial Cells from Subcutaneous Fat Tissue Ideal for Direct Cell Seeding

Cited by 13 publications

References 6 publications

Readily Available Tissue-Engineered Vascular Grafts

Readily Available Tissue-Engineered Vascular Grafts

MicroRNA-181b Improves Glucose Homeostasis and Insulin Sensitivity by Regulating Endothelial Function in White Adipose Tissue

Extraction of cells for single‐stage seeding of vascular‐bypass grafts

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