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Cell sheet engineering using the stromal vascular fraction of adipose tissue as a vascularization strategy

机译:使用脂肪组织的基质血管分数作为血管形成策略的细胞片工程

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摘要

Current vascularization strategies for Tissue Engineering constructs, in particular cell sheet-based, are limited by time-consuming and expensive endothelial cell isolation and/or by the complexity of using extrinsic growth factors. Herein, we propose an alternative strategy using angiogenic cell sheets (CS) obtained from the stromal vascular fraction (SVF) of adipose tissue that can be incorporated into more complex constructs. Cells from the SVF were cultured in normoxic and hypoxic conditions for up to 8 days in the absence of extrinsic growth factors. Immunocytochemistry against CD31 and CD146 revealed spontaneous organization in capillary-like structures, more complex after hypoxic conditioning. Inhibition of HIF-1 alpha pathway hindered capillary-like structure formation in SVF cells cultured in hypoxia, suggesting a role of HIF-1 alpha. Moreover, hypoxic SVF cells showed a trend for increased secretion of angiogenic factors, which was reflected in increased network formation by endothelial cells cultured on matrigel using that conditioned medium. In vivo implantation of SVF CS in a mouse hind limb ischemia model revealed that hypoxia-conditioned CS led to improved restoration of blood flow. Both in vitro and in vivo data suggest that SVF CS can be used as simple and cost-efficient tools to promote functional vascularization of TE constructs.
机译:用于组织工程构建体的当前血管化策略,特别是细胞片基础,受耗时和昂贵的内皮细胞分离和/或使用外本生长因子的复杂性受到限制。在此,我们提出了使用从脂肪组织的基质血管级分(SVF)获得的血管生成细胞片(CS)提出了可以掺入更复杂的构建体中的抗血管生成细胞片(CS)。从SVF的细胞在常氧和缺氧条件下培养,在没有外部生长因子的情况下长达8天。针对CD31和CD146的免疫细胞化学揭示了毛细血管状结构的自发组织,缺氧调理后更复杂。抑制HIF-1α途径阻碍缺氧培养的SVF细胞中的毛细管样结构形成,表明HIF-1α的作用。此外,缺氧SVF细胞显示出增加血管生成因子的分泌增加的趋势,这在使用该条件培养基的基质胶中培养的内皮细胞增加了网络形成。在小鼠后肢缺血模型中植入SVF CS的SVF CS揭示了缺氧调节的CS导致血流恢复。体外和体内数据都表明SVF CS可以用作简单且具有成本效益的工具,以促进TE构建体的功能血管化。

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