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脱细胞肝素包被血管移植物的碱性成纤维细胞生长因子包被及内皮细胞接种

Basic fibroblast growth factor coating and endothelial cell seeding of a decellularized heparin-coated vascular graft.

作者信息

Conklin Brian S, Wu Huakang, Lin Peter H, Lumsden Alan B, Chen Changyi

机构信息

Molecular Surgeon Research Center, Division of Vascular Surgery and Endovascular Therapy, Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX 77030, USA.

出版信息

Artif Organs. 2004 Jul;28(7):668-75. doi: 10.1111/j.1525-1594.2004.00062.x.

Abstract

The objective of this study was to determine the effect of basic fibroblast growth factor (bFGF) coating on endothelial cell seeding and proliferation on a decellularized heparin coated vascular graft and to determine the retention of seeded cells on the graft under flow conditions. Disks of heparin coated decellularized grafts were incubated for 24 h as controls or with bFGF. Human microvascular endothelial cells (HMECs) or canine peripheral blood endothelial progenitor cells (CEPC) were seeded onto the disks and incubated for 96 h or 48 h, respectively. HMECs were also seeded onto the luminal surfaces of two heparin-coated decellularized grafts for 3 h. One graft was placed in a perfusion culture system and cultured for an additional 6 h with flow and pressure. After culturing, there were 4.7 +/- 1.4 cells/mm(2) HMECs on control grafts and 11.4 +/- 1.4 cells/mm(2) in bFGF treated grafts (P < 0.05). Likewise, with CEPCs, there were 14.8 +/- 4.8 cells/mm(2) in control grafts and 33.3 +/- 7.3 cells/mm(2) in bFGF treated grafts. After only 3 h of cell attachment, 60% of HMECs were retained in the intact graft exposed flow relative to the static control graft, which is an acceptable level. These data demonstrate that bFGF coating on the heparin bound decellularized grafts significantly increases both HMEC and dog EPC proliferation and that seeded cells are stable under perfusion conditions.

摘要

本研究的目的是确定碱性成纤维细胞生长因子(bFGF)包被对去细胞肝素包被血管移植物上内皮细胞接种和增殖的影响,并确定在流动条件下接种细胞在移植物上的留存情况。将肝素包被的去细胞移植物圆盘作为对照或与bFGF一起孵育24小时。将人微血管内皮细胞(HMECs)或犬外周血内皮祖细胞(CEPC)接种到圆盘上,分别孵育96小时或48小时。HMECs也接种到两个肝素包被的去细胞移植物的管腔表面3小时。将一个移植物置于灌注培养系统中,在有流动和压力的情况下再培养6小时。培养后,对照移植物上有4.7±1.4个细胞/mm²的HMECs,bFGF处理的移植物上有11.4±1.4个细胞/mm²(P<0.05)。同样,对于CEPCs,对照移植物上有14.8±4.8个细胞/mm²,bFGF处理的移植物上有33.3±7.3个细胞/mm²。仅细胞附着3小时后,相对于静态对照移植物,60%的HMECs留存于完整的暴露于流动的移植物中,这是一个可接受的水平。这些数据表明,肝素结合的去细胞移植物上的bFGF包被显著增加了HMEC和犬EPC的增殖,并且接种的细胞在灌注条件下是稳定的。

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