Bérard Xavier, Rémy-Zolghadri Murielle, Bourget Chantal, Turner Neill, Bareille Reine, Daculsi Richard, Bordenave Laurence
INSERM, U577, Bordeaux and Université Victor Segalen Bordeaux 2, UMR-577, Bordeaux F-33076, France.
Acta Biomater. 2009 May;5(4):1147-57. doi: 10.1016/j.actbio.2008.10.002. Epub 2008 Oct 17.
One of the goals of vascular tissue engineering is to create functional conduits for small-diameter bypass grafting. The present biocompatibility study was undertaken to check the ability of cord blood progenitor-derived endothelial cells (PDECs) to take the place of endothelial cells in vascular tissue engineering. After isolation, culture and characterization of endothelial progenitor cells, the following parameters were explored, with a commercial knitted polyester prosthesis (Polymaille C, Laboratoires Pérouse, France) impregnated with collagen: cell adhesion and proliferation, colonization, cell retention on exposure to flow, and the ability of PDECs to be regulated by arterial shear stress via mRNA levels. PDECs were able to adhere to commercial collagen-coated vascular grafts in serum-free conditions, and were maintained but did not proliferate when seeded at 2.0 x 10(5) cm(-2). Cellularized conduits were analyzed by histology and histochemical staining, demonstrating collagen impregnation and the endothelial characteristics of the colonizing cells. Thirty-six hours after cell seeding the grafts were maintained for 6 h of either static conditions (controls) or application of pulsatile laminar shear stress, which restored the integrity of the monolayer. Finally, quantitative real-time RT-PCR analysis performed at 4 and 8 h from cells lining grafts showed that MMP1 mRNA only was increased at 4h whereas vWF, VE-cadherin and KDR were not significantly modified at 4 and 8 h. Our results show that human cord blood PDECs are capable of forming an efficient lining and to withstand shear stress.
血管组织工程的目标之一是创建用于小直径旁路移植的功能性管道。本生物相容性研究旨在检验脐带血祖细胞衍生的内皮细胞（PDEC）在血管组织工程中替代内皮细胞的能力。在对内皮祖细胞进行分离、培养和鉴定后，对以下参数进行了探索：使用浸渍有胶原蛋白的商用针织聚酯假体（法国佩鲁斯实验室的Polymaille C），研究细胞黏附与增殖、定植、暴露于流动状态下的细胞保留情况，以及PDEC通过mRNA水平受动脉剪切应力调节的能力。PDEC能够在无血清条件下黏附于商用胶原蛋白包被的血管移植物，当以2.0×10⁵个/cm²接种时可维持存活但不增殖。通过组织学和组织化学染色分析细胞化管道，证实了胶原蛋白浸渍以及定植细胞的内皮特征。细胞接种36小时后，将移植物在静态条件（对照）或施加脉动层流剪切应力下维持6小时，结果恢复了单层的完整性。最后，对移植物内衬细胞在4小时和8小时进行的定量实时RT-PCR分析表明，仅MMP1 mRNA在4小时时增加，而vWF、VE-钙黏蛋白和KDR在4小时和8小时时未发生显著改变。我们的结果表明，人脐带血PDEC能够形成有效的内衬并承受剪切应力。
