Cell sheet engineering using the stromal vascular fraction of adipose tissue as a vascularization strategy
Cell sheet engineering using the stromal vascular fraction of adipose tissue as a vascularization strategy
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DOI：
10.1016/j.actbio.2017.03.034
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发表时间：
2017-06-01
影响因子：
9.7
通讯作者：
Reis, Rui L.
中科院分区：
文献类型：
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作者：
Costa, Marina;Cerqueira, Mariana T.;Reis, Rui L.
目前用于组织工程构建体的血管化策略，特别是基于细胞片的血管化策略，受到耗时且昂贵的内皮细胞分离和/或使用外源性生长因子的复杂性的限制。在此，我们提出了一种替代策略，使用血管生成细胞片（CS）从基质血管部分（SVF）的脂肪组织，可以纳入到更复杂的结构。在不存在外源性生长因子的情况下，将来自SVF的细胞在常氧和低氧条件下培养长达8天。对CD 31和CD 146的免疫细胞化学显示自发组织的毛细血管样结构，更复杂的缺氧条件。HIF-1 α通路的抑制阻碍了缺氧培养的SVF细胞中毛细血管样结构的形成，这表明HIF-1 α的作用。此外，缺氧SVF细胞显示出血管生成因子分泌增加的趋势，这反映在使用该条件培养基在基质胶上培养的内皮细胞的网络形成增加。在小鼠后肢缺血模型中体内植入SVF CS显示，低氧条件CS导致血流恢复改善。无论是在体外和体内的数据表明，SVF CS可以被用来作为简单和成本效益的工具，以促进TE constructs.Statement of SignificanceNeovascularization的功能血管化植入后是一个主要的障碍，为生产临床上可行的细胞片为基础的组织工程constructs. Statement的。使用内皮细胞和外源性血管生成生长因子的策略是昂贵和耗时的，并且可能引起致瘤性的关注。在这份手稿中，我们描述了一种简化的方法，使用血管生成细胞片制造的基质血管部分的脂肪组织。这些细胞片层的强血管生成行为，在不使用外部生长因子的情况下实现，通过低氧培养进一步刺激。当植入到后肢缺血的体内模型中时，血管生成细胞片有助于血流量恢复。因此，这些细胞片可用作增加基于细胞片的厚构建体的新血管形成的直接工具。(C)2017由Elsevier Ltd代表Acta Materialia Inc.发布。
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.Statement of SignificanceNeovascularization after implantation is a major obstacle for producing clinically viable cell sheet-based tissue engineered constructs. Strategies using endothelial cells and extrinsic angiogenic growth factors are expensive and time consuming and may raise concerns of tumorigenicity. In this manuscript, we describe a simplified approach using angiogenic cell sheets fabricated from the stromal vascular fraction of adipose tissue. The strong angiogenic behavior of these cell sheets, achieved without the use of external growth factors, was further stimulated by low oxygen culture. When implanted in an in vivo model of hind limb ischemia, the angiogenic cell sheets contributed to blood flux recovery. These cell sheets can therefore be used as a straightforward tool to increase the neovascularization of cell sheet-based thick constructs. (C) 2017 Published by Elsevier Ltd on behalf of Acta Materialia Inc.